Immunotherapy of cancer

ABSTRACT

Immunogenic modulators and compositions comprising oligonucleotide agents capable of inhibiting suppression of immune response by reducing expression of one or more gene involved with an immune response suppression mechanism.

CROSS REFERENCE

This application claims priority to U.S. Provisional Application No. 61/910,728, filed Dec. 2, 2013, herein incorporated by reference in its entirety.

TECHNICAL FIELD

The present invention relates to immunogenic compositions, method of making immunogenic compositions, and methods of using immunogenic compositions for the treatment of cell proliferative disorders or infectious disease, including, for example, cancer and autoimmune disorders.

More particularly, the invention provides cells that are treated with oligonucleotides specifically designed to modulate expression of target genes involved in tumor immune resistance mechanisms.

BACKGROUND

Immunotherapy is the “treatment of disease by inducing, enhancing, or suppressing an immune response”. Immunotherapies designed to elicit or amplify an immune response are activation immunotherapies, while immunotherapies that reduce or suppress immune response are classified as suppression immunotherapies.

Immunotherapy of cancer has become increasingly important in clinical practice over recent decades. The primary approach in today's standard of care is passive immunotherapy through the use of recombinant monoclonal antibodies (mAbs). MAbs act through a mechanisms relevant to the body's own humoral immune response, by binding to key antigens involved in the tumor development and causing moderate forms of cell-mediated immunity, such as antibody-dependent cell-mediated cytotoxicity (ADCC).

Another group of emerging immunotherapeutic approaches is based on the administration of cells capable of destroying tumor cells. The administered cells may be the patient's own tumor-infiltrating lymphocytes (TIL), isolated and expanded ex-vivo. In some cases, TIL are capable of recognizing a variety of tumor associated antigens (TAA), while in other cases TIL can be reactivated and expanded in vitro to recognize specific antigens. The TIL-based therapeutic approaches are commonly referred to as “adoptive cell transfer” (ACT).

Further developments of ACT involve genetic modifications of T-cells to express receptors that recognize specific tumor-associated antigens (TAA). Such modifications may induce the expression of a specific T-cell receptor (TCR) or of a chimeric antigen receptor (CAR) consisting of TAA-specific antibody fused to CD3/co-stimulatory molecule transmembrane and cytoplasmic domains.

The ACT methods may also be considered as passive immunotherapeutic approaches in that they act directly on the tumor cells without invoking an extended immune response. However, unlike mAbs, ACT agents are capable of fully destroying the tumor cells, as opposed to the blockade of selected receptors and moderate cellular responses such as ADCC.

There is ongoing development of numerous methods of active immunotherapy, which restore the ability of body's own immune system to generate antitumor response. Active immunotherapeutic agents are often called therapeutic cancer vaccines, or just cancer vaccines. Many cancer vaccines are currently in clinical trials, and sipuleucell-T has recently become the first such vaccine approved by the United States FDA.

There are several classes of cancer vaccines using different antigens and different mechanisms of generating cell-mediated immune response. One class of vaccines is based on peptide fragments of antigens selectively expressed by tumor cells. The peptides are administered alone or in combination with immune-stimulatory agents, which may include adjuvants and cytokines, such as granulocyte-macrophage colony-stimulating factor (GM-CSF).

Another class of cancer vaccines is based on modified (e.g. sub-lethally irradiated) tumor cells used as antigens, also in combinations with immunostimulatory agents. Vaccines of this type currently in clinical trials are based both on autologous (e.g. OncoVAX, LipoNova) and allogeneic (e.g. Canvaxin, Onyvax-P, GVAX) tumor cell lines.

Yet another class of cancer vaccines uses dendritic cells. By their nature, dendritic cells (DC) are “professional” antigen-presenting cells capable of generating of a strong antigen-dependent cell-mediated immune response and eliciting therapeutic T-cells in vivo. DC-based cancer vaccines usually comprise DCs isolated from patients or generated ex vivo by culturing patient's hematopoietic progenitor cells or monocytes. DCs are further loaded with tumor antigens and sometimes combined with immune-stimulating agents, such as GM-CSF. A large number of DC-vaccines are now in clinical trials, and the first FDA-approved vaccine sipuleucell-T is based on DC.

Mechanisms of Immunosuppression and Therapeutic Approaches to its Mitigation

One of the key physiologic functions of the immune system is to recognize and eliminate neoplastic cells, therefore an essential part of any tumor progression is the development of immune resistance mechanisms. Once developed, these mechanisms not only prevent the natural immune system from effecting the tumor growth, but also limit the efficacy of any immunotherapeutic approaches to cancer. An important immune resistance mechanism involves immune-inhibitory pathways, sometimes referred to as immune checkpoints. The immune-inhibitory pathways play particularly important role in the interaction between tumor cells and CD8+ cytotoxic T-lymphocytes, including ACT therapeutic agents. Among important immune checkpoints are inhibitory receptors expressed on the T-cell surface, such as CTLA-4, PD1 and LAG3, among others.

The importance of the attenuation of immune checkpoints has been recognized by the scientific and medical community. One way to mitigate immunosuppression is to block the immune checkpoints by specially designed agents. The CTLA-4-blocking-antibody, ipilimumab, has recently been approved by the FDA. Several molecules blocking PD1 are currently in clinical development.

Immunosuppression mechanisms also negatively affect the function of dendritic cells and, as a consequence, the efficacy of DC-based cancer vaccines. Immunosuppressive mechanisms can inhibit the ability of DC to present tumor antigens through the MHC class I pathway and to prime naïve CD8+ T-cells for antitumor immunity. Among the important molecules responsible for the immunosuppression mechanisms in DC are ubiquitin ligase A20 and the broadly immune-suppressive protein SOCS1.

The efficacy of immunotherapeutic approaches to cancer can be augmented by combining them with inhibitors of immune checkpoints. Numerous ongoing preclinical and clinical studies are exploring potential synergies between cancer vaccines and other immunotherapeutic agents and checkpoint blocking agents, for example, ipilimumab. Such combination approaches have the potential to result in significantly improved clinical outcomes.

However, there are a number of drawbacks of using cancer immunotherapeutic agents in combination with checkpoint inhibitors. For example, immune checkpoint blockade can lead to the breaking of immune self-tolerance, thereby inducing a novel syndrome of autoimmune/auto-inflammatory side effects, designated “immune related adverse events,” mainly including rash, colitis, hepatitis and endocrinopathies (Corsello, et al. J. Clin. Endocrinol. Metab., 2013, 98:1361).

Reported toxicity profiles of checkpoint inhibitors are different than the toxicity profiles reported for other classes of oncologic agents. Those involve inflammatory events in multiple organ systems, including skin, gastrointestinal, endocrine, pulmonary, hepatic, ocular, and nervous system. (Hodi, 2013, Annals of Oncology, 24: Suppl, i7).

In view of the above, there is a need for new cancer therapeutic agents that can be used in combination with checkpoint inhibitors as well as other classes of oncolytic agents without risk of adverse inflammatory events in multiple organ systems previously reported for checkpoint inhibitors. The immunotherapeutic cells of the invention, prepared by treating cells with a combination oligonucleotide agents targeting genes associated with tumor or infections disease resistance mechanisms, as well as methods of producing such therapeutic cells and methods of treating disease with the produced therapeutic cells, satisfy this long felt need.

SUMMARY OF EMBODIMENTS OF THE INVENTION

The efficacy of immunotherapeutic approaches to cell proliferation disorders and infectious diseases can be augmented by combining them with inhibitors of immune checkpoints. Numerous synergies between cancer vaccines and other immunotherapeutic agents and checkpoint blocking agents provide opportunities for combination approaches that may significantly improve clinical outcomes for example, in proliferative cell disorders and immune diseases.

Various embodiments of the inventions disclosed herein include compositions comprising therapeutic cells obtained by treating cells ex vivo with oligonucleotides to modulate expression of target genes involved in immune suppression mechanisms. The oligonucleotide agent may be an antisense oliogonucleotide (ASO), including locked nucleic acids (LNAs), methoxyethyl gapmers, and the like, or an siRNA, miRNA, miRNA-inhibitor, morpholino, PNA, and the like. The oligonucleotide is preferably a self-delivered (sd) RNAi agent. The oligonucleotides may be chemically modified, for example, including at least one 2-O-methyl modification, 2′-Fluro modification, and/or phosphorothioate modification. The oligonucleotides may include one or more hydrophobic modification, for example, one or more sterol, cholesterol, vitamin D, Naphtyl, isobutyl, benzyl, indol, tryptophane, or phenyl hydrophobic modification. The oligonucleotide may be a hydrophobically-modified siRNA-antisense hybrid. The oligonucleotides may be used in combination with transmembrane delivery systems, such as delivery systems comprising lipids.

In an embodiment, the cells are obtained and/or derived from a cancer or infectious disease patient, and may be, for example, tumor infiltrating lymphocytes (TIL) and/or T-cells, antigen presenting cells such as dendritic cells, natural killer cells, induced-pluripotent stem cells, stem central memory T-cells, and the like. The T-cells and NK-cells are preferably genetically engineered to express high-affinity T-Cell receptors (TCR) and/or chimeric antibody or antibody-fragment—T-Cell receptors (CAR). In an embodiment, the chimeric antibody/antibody fragment is preferably capable of binding to antigens expressed on tumor cells. Immune cells may be engineered by transfection with plasmid, viral delivery vehicles, or mRNAs.

In an embodiment, the chimeric antibody or fragment is capable of binding CD19 receptors of B-cells and/or binding to antigens expressed on tumors, such as melanoma tumors. Such melanoma-expressed antigens include, for example, GD2, GD3, HMW-MAA, VEGF-R2, and the like.

Target genes identified herein for modification include: cytotoxic T-cell antigen 4 (CTLA4), programmed cell death protein 1 (PD1), tumor growth factor receptor beta (TGFR-beta), LAG3, TIM3, and adenosine A2a receptor; anti-apoptotic genes including, but not limited to: BAX, BAC, Casp8, and P53; A20 ubiquitine ligase (TNFAIP3, SOCS1 (suppressor of cytokine signaling), IDO (indolamine-2,3-dioxygenase; tryptophan-degrading enzyme), PD-L1 (CD274)(surface receptor, binder to PD1 on Tcells), Notch ligand Deltal (DLL1), Jagged 1, Jagged 2, FasL (pro-apoptotic surface molecule), CCL17, CCL22 (secreted chemokines that attract Treg cells), IL10 receptor (IL10RA), p38 (MAPK14), STAT3, TNFSF4 (OX40L), MicroRNA miR-155, miR-146a, anti-apoptotic genes including but not limited to BAX, BAC, Casp8 and P53, and the like genes, and combinations thereof. Representative target sequences are listed in Table 1.

The engineered therapeutic cells are treated with RNAi agents designed to inhibit expression of one or more of the targeted genes. The RNAi agent may comprise a guide sequence that hybridizes to a target gene and inhibits expression of the target gene through an RNA interference mechanism, where the target region is selected from the group listed in Table 1. The RNA agent can be chemically modified, and preferably includes at least one 2′-O-methyl, 2′-O-Fluoro, and/or phosphorothioate modification, as well as at least one hydrophobic modification such as cholesterol, and the like.

The immunogenic compositions described herein are useful for the treatment of proliferative disorders, including cancers, and/or infectious disease and are produced by the ex-vivo treatment of cells with oligonucleotides to modulate the expression of target genes involved in tumor immune resistance mechanisms. The ex vivo treatment of cells includes administering to the cells an oligonucleotide capable of targeting and inhibiting expression of a gene involved in a tumor suppressor mechanism, such as the genes listed in Table 1. The oligonucleotide can be used in combination with a transmembrane delivery system that may comprise one or more of: lipid(s) and vector, such as a viral vector.

The invention includes a method of treating a cell proliferative disorder or infectious disease by administering to a subject in need thereof, an immunogenic composition comprising cells that have been treated with one or more oligonucleotide to modulate the expression of one or more target gene involved in tumor immune resistance mechanisms, for example, one or more of the target genes of Table 1.

The invention preferably includes immunogenic cells treated with a plurality of oligonucleotide agents targeting a combination of target genes described herein. The combination may target a plurality of suppressor receptor genes, cytokine receptor genes, regulatory genes, and/or apoptotic factors in order to inhibit tumor immune resistance mechanisms.

The present invention is directed to novel immunotherapeutic cells, methods of generating the immunotherapeutic cells, and therapeutic methods employing such cells.

A new method of immune checkpoint inhibition is described herein, applicable to a broad variety of cell-based immunotherapies, including, but not limited to adaptive cell transfer, for example, based on TIL, TCR, CAR, and other cell types, as well as dendritic cell-based cancer vaccines. Self-deliverable RNAi technology provides efficient transfection of short oligonucleotides in any cell type, including immune cells, providing increased efficacy of immunotherapeutic treatments. In addition, the activated immune cells can be protected by preventing apoptosis via inhibition of key activators of the apoptotic pathway, such as BAC, BAX, Casp8, and P53, among others.

The activated immune cells modified by oligonucleotide transfer for a single therapeutic agent for administration to a subject, providing a number of advantages as compared to separately administered combinations of vaccines and immunotherapeutics and separately administered checkpoint inhibitors. These advantages include lack of side effects associated with the checkpoint inhibitors in a single therapeutic agent (activated immune cells modified by oligonucleotides targeting immune resistance genes).

The claimed immunotherapeutic cells, method of producing immunotherapeutic cells by introduction of oligonucleotide molecules targeting immune resistance pathways, and methods of treating proliferative and infectious disease, improves upon any known immunotherapeutic cells and methods of producing immunotherapeutic cells because it provides:

-   -   1) a single therapeutic composition providing a combination of         checkpoint inhibitors and other immune resistance mechanism         inhibitors;     -   2) with reduced toxicity; and     -   3) increased efficacy as compared with other compositions.

BRIEF DESCRIPTION OF THE FIGURES

The foregoing features of embodiments will be more readily understood by reference to the following detailed description, taken with reference to the accompanying drawings, in which:

FIG. 1 is a schematic diagram showing the structure of an sdRNA molecule.

FIG. 2 is a graph showing sdRNA-induced silencing of GAPDH and MAP4K4 in HeLa cells.

FIG. 3 is a graph showing sdRNA-induced knock-down of multiple targets using sdRNA agents directed to three genes in NK-92 cells.

FIG. 4 is a graph showing the knock-down of gene expression in Human Primary T cells by sdRNA agents targeting TP53 and MAP4K4.

FIG. 5 is a graph showing sdRNA-induced knock-down of CTLA4 and PD1 in Human Primary T cells.

FIG. 6 is a graph showing the reduction of PDCD1 and CTLA-4 surface expression by sdRNA in Human Primary T cells.

FIG. 7 is a graph showing MAP4K4-cy3 sdRNA delivery into T and B cells in human PBMCs.

DETAILED DESCRIPTION OF SPECIFIC EMBODIMENTS

The invention is defined by the claims, and includes oligonucleotides specifically designed and selected to reduce and/or inhibit expression of suppressors of immune resistance (inhibitory oligonucleotides), compositions comprising cells modified by treatment with such inhibitory oligonucleotides, methods of making such compositions, and methods of using the compositions to treat proliferation and/or infectious diseases. In particular, cells are treated with a combination of oligonucleotide agents, each agent particularly designed to interfere with and reduce the activity of a targeted immune suppressor.

Preferably, the combination of oligonucleotide agents targets multiple immune suppressor genes selected from checkpoint inhibitor genes such as CTLA4, PD-1/PD-1L, BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3, B7-H4 receptors, and TGF beta type 2 receptor; cytokine receptors that inactivate immune cells, such as TGF-beta receptor A and IL-10 receptor; regulatory genes/transcription factors modulating cytokine production by immune cells, such as STAT-3 and P38, miR-155, miR-146a; and apoptotic factors involved in cascades leading to cell death, such as p53 and Cacp8.

Most preferably the oligonucleotide agent is a self-deliverable RNAi agent, which is a hydrophobically modified siRNA-antisense hybrid molecule, comprising a double-stranded region of about 13-22 base pairs, with or without a 3′-overhang on each of the sense and antisense strands, and a 3′ single-stranded tail on the antisense strand of about 2-9 nucleotides. The oligonucleotide contains at least one 2′-O-Methyl modification, at least one 2′-O-Fluoro modification, and at least one phosphorothioate modification, as well as at least one hydrophobic modification selected from sterol, cholesterol, vitamin D, napthyl, isobutyl, benzyl, indol, tryptophane, phenyl, and the like hydrophobic modifiers (see FIG. 1). The oligonucleotide may contain a plurality of such modifications.

Definitions

As used in this description and the accompanying claims, the following terms shall have the meanings indicated, unless the context requires otherwise:

Proliferative disease, as used herein, includes diseases and disorders characterized by excessive proliferation of cells and turnover of cellular matrix, including cancer, atherlorosclerosis, rheumatoid arthritis, psoriasis, idiopathic pulmonary fibrosis, scleroderma, cirrhosis of the liver, and the like. Cancers include but are not limited to, one or more of: small cell lung cancer, colon cancer, breast cancer, lung cancer, prostate cancer, ovarian cancer, pancreatic cancer, melanoma, hematological malignancy such as chronic myeloid leukemia, and the like cancers where immunotherapeutic intervention to suppress tumor related immune resistance is needed.

Immune target genes can be grouped into at least four general categories: (1) checkpoint inhibitors; (2) cytokine receptors that inactivate immune cells, (3) anti-apoptotic genes; and (4) regulator genes, for example, transcription factors.

Immune Checkpoint inhibitors (ICI), as used herein, include immunotherapeutic agents that bind to certain checkpoint proteins, such as cytotoxic T lymphocyte antigen-4 (CTLA-4) and programmed death-1 (PD-1) and its ligand PD-L1 to block and disable inhibitory proteins that prevent the immune system from attacking diseased cells such as cancer cells, liberating tumor-specific T cells to exert their effector function against tumor cells.

Tumor related immune resistance genes, as used herein, include genes involved in checkpoint inhibition of immune response, such as CTLA-4 and PD-1/PD-L1; TGF-beta, LAG3, Tim3, adenosine A2a receptor;

Regulator genes, as used herein, include transcription factors and the like that modulate cytokine production by immune cells, and include p38, STAT3, microRNAs miR-155, miR-146a;

Anti-apoptotic genes, as used herein, include BAX, BAC, Casp8, P53 and the like; and combinations thereof.

Infectious diseases, as used herein, include, but are not limited to, diseases caused by pathogenic microorganisms, including, but not limited to, one or more of bacteria, viruses, parasites, or fungi, where immunotherapeutic intervention to suppress pathogen related immune resistance and/or overactive immune response.

Immunogenic composition, as used herein, includes cells treated with one or more oligonucleotide agent, wherein the cells comprise T-cells. The T-cells may be genetically engineered, for example, to express high affinity T-cell receptors (TCR), chimeric antibody—T-cell receptors (CAR), where the chimeric antibody fragments are capable of binding to CD19 receptors of B-cells and/or to antigens expressed on tumor cells. In one embodiment, the chimeric antibody fragments bind antigens expressed on melanoma tumors, selected from GD2, GD3, HMW-MAA, and VEGF-R2.

Immunogenic compositions described herein include cells comprising antigen-presenting cells, dendritic cells, engineered T-cells, natural killer cells, stem cells, including induced pluripotent stem cells, and stem central memory T-cells. The treated cell also comprises one or a plurality of oligonucleotide agents, preferably sdRNAi agents specifically targeting a gene involved in an immune suppression mechanism, where the oligonucleotide agent inhibits expression of said target gene.

In one embodiment, the target gene is selected from A20 ubiquitin ligase such as TNFAIP3, SOCS1 (suppressor of cytokine signaling), Tyro3/Axl/Mer (suppressors of TLR signaling), IDO (indolamine-2,3-dioxygenase, tryptophan-degrading enzyme), PD-L1/CD274 (surface receptor, binds PD1 on T-cells), Notch ligand Delta (DLL1), Jagged 1, Jagged 2, FasL (pro-apoptotic surface molecule), CCL17, CCL22 (secreted chemokines that attract Treg cells), IL-10 receptor (IL10Ra), p38 (MAPK14), STAT3, TNFSF4 (OX40L), microRNA miR-155, miR-146a, anti-apoptotic genes, including but not limited to BAX, BAC, Casp8, and P53; and combinations thereof.

Particularly preferred target genes are those shown in Table 1.

Ex-vivo treatment, as used herein, includes cells treated with oligonucleotide agents that modulate expression of target genes involved in immune suppression mechanisms. The oligonucleotide agent may be an antisense oligonucleotide, including, for example, locked nucleotide analogs, methyoxyethyl gapmers, cyclo-ethyl-B nucleic acids, siRNAs, miRNAs, miRNA inhibitors, morpholinos, PNAs, and the like. Preferably, the oligonucleotide agent is an sdRNAi agent targeting a gene involved in an immune suppression mechanism. The cells treated in vitro by the oligonucleotide agent may be immune cells expanded in vitro, and can be cells obtained from a subject having a proliferative or infectious disease. Alternatively, the cells or tissue may be treated in vivo, for example by in situ injection and/or intravenous injection.

Oligonucleotide or oligonucleotide agent, as used herein, refers to a molecule containing a plurality of “nucleotides” including deoxyribonucleotides, ribonucleotides, or modified nucleotides, and polymers thereof in single- or double-stranded form. The term encompasses nucleotides containing known nucleotide analogs or modified backbone residues or linkages, which are synthetic, naturally occurring, and non-naturally occurring, which have similar binding properties as the reference nucleic acid, and which are metabolized in a manner similar to the reference nucleotides. Examples of such analogs include, without limitation, phosphorothioates, phosphoramidates, methyl phosphonates, chiral-methyl phosphonates, 2-O-methyl ribonucleotides, peptide-nucleic acids (PNAs).

Nucleotide, as used herein to include those with natural bases (standard), and modified bases well known in the art. The nucleotides can be unmodified or modified at the sugar, phosphate and/or base moiety, (also referred to interchangeably as nucleotide analogs, modified nucleotides, non-natural nucleotides, non-standard nucleotides and other; see, for example, PCT Publications No. WO 92/07065 and WO 93/15187. Non-limiting examples of base modifications that can be introduced into nucleic acid molecules include, hypoxanthine, purine, pyridin-4-one, pyridin-2-one, phenyl, pseudouracil, 2,4,6-trimethoxy benzene, 3-methyl uracil, dihydrouridine, naphthyl, aminophenyl, 5-alkylcytidines (e.g., 5-methylcytidine), 5-alkyluridines (e.g., ribothymidine), 5-halouridine (e.g., 5-bromouridine) or 6-azapyrimidines or 6-alkylpyrimidines (e.g. 6-methyluridine and pseudouridine), propyne, and others. The phrase “modified bases” includes nucleotide bases other than adenine, guanine, cytosine, and uracil, modified for example, at the 1′ position or their equivalents.

As used herein, the term “deoxyribonucleotide” encompasses natural and synthetic, unmodified and modified deoxyribonucleotides. Modifications include changes to the sugar moiety, to the base moiety and/or to the linkages between deoxyribonucleotide in the oligonucleotide.

As used herein, the term “RNA” defines a molecule comprising at least one ribonucleotide residue. The term “ribonucleotide” defines a nucleotide with a hydroxyl group at the 2′ position of a □-D-ribofuranose moiety. The term RNA includes double-stranded RNA, single-stranded RNA, isolated RNA such as partially purified RNA, essentially pure RNA, synthetic RNA, recombinantly produced RNA, as well as altered RNA that differs from naturally occurring RNA by the addition, deletion, substitution and/or alteration of one or more nucleotides. Nucleotides of the RNA molecules described herein may also comprise non-standard nucleotides, such as non-naturally occurring nucleotides or chemically synthesized nucleotides or deoxynucleotides. These altered RNAs can be referred to as analogs or analogs of naturally-occurring RNA.

As used herein, “modified nucleotide” refers to a nucleotide that has one or more modifications to the nucleoside, the nucleobase, pentose ring, or phosphate group. For example, modified nucleotides exclude ribonucleotides containing adenosine monophosphate, guanosine monophosphate, uridine monophosphate, and cytidine monophosphate and deoxyribonucleotides containing deoxyadenosine monophosphate, deoxyguanosine monophosphate, deoxythymidine monophosphate, and deoxycytidine monophosphate. Modifications include those naturally-occurring that result from modification by enzymes that modify nucleotides, such as methyltransferases.

Modified nucleotides also include synthetic or non-naturally occurring nucleotides. Synthetic or non-naturally occurring modifications in nucleotides include those with 2′ modifications, e.g., 2′-O-methyl, 2′-methoxyethoxy, 2′-fluoro, 2′-allyl, 2′-O-[2-(methylamino)-2-oxoethyl], 4′-thio, 4′-CH2-O-2′-bridge, 4′-(CH2) 2-O-2′-bridge, 2′-LNA, and 2′-O-(N-methylcarbamate) or those comprising base analogs. In connection with 2′-modified nucleotides as described for the present disclosure, by “amino” is meant 2′-NH2 or 2′-O—NH2, which can be modified or unmodified. Such modified groups are described, for example, in U.S. Pat. Nos. 5,672,695 and 6,248,878.

As used herein, “microRNA” or “miRNA” refers to a nucleic acid that forms a single-stranded RNA, which single-stranded RNA has the ability to alter the expression (reduce or inhibit expression; modulate expression; directly or indirectly enhance expression) of a gene or target gene when the miRNA is expressed in the same cell as the gene or target gene. In one embodiment, a miRNA refers to a nucleic acid that has substantial or complete identity to a target gene and forms a single-stranded miRNA. In some embodiments miRNA may be in the form of pre-miRNA, wherein the pre-miRNA is double-stranded RNA. The sequence of the miRNA can correspond to the full length target gene, or a subsequence thereof. Typically, the miRNA is at least about 15-50 nucleotides in length (e.g., each sequence of the single-stranded miRNA is 15-50 nucleotides in length, and the double stranded pre-miRNA is about 15-50 base pairs in length). In some embodiments the miRNA is 20-30 base nucleotides. In some embodiments the miRNA is 20-25 nucleotides in length. In some embodiments the miRNA is 20, 21, 22, 23, 24, 25, 26, 27, 28, 29, or 30 nucleotides in length.

Target gene, as used herein, includes genes known or identified as modulating the expression of a gene involved in an immune resistance mechanism, and can be one of several groups of genes, such as suppressor receptors, for example, CTLA4 and PD1; cytokine receptors that inactivate immune cells, for example, TGF-beta receptor, LAG3, Tim3, adenosine A2a receptor, and IL10 receptor; regulatory genes for example, STAT3, p38, mir155 and mir146a; and apoptosis factors involved in cascades leading to cell death, for example, P53, Casp8, BAX, BAC, and combinations thereof. See also preferred target genes listed in Table 1.

As used herein, small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, defines a group of double-stranded RNA molecules, comprising sense and antisense RNA strands, each generally of about 1022 nucletides in length, optionally including a 3′ overhang of 1-3 nucleotides. siRNA is active in the RNA interference (RNAi) pathway, and interferes with expression of specific target genes with complementary nucleotide sequences.

As used herein, sdRNA refers to “self-deliverable” RNAi agents, that are formed as an asymmetric double-stranded RNA-antisense oligonucleotide hybrid. The double stranded RNA includes a guide (sense) strand of about 19-25 nucleotides and a passenger (antisense) strand of about 10-19 nucleotides with a duplex formation that results in a single-stranded phosphorothiolated tail of about 5-9 nucleotides.

The RNA sequences may be modified with stabilizing and hydrophobic modifications such as sterols, for example, cholesterol, vitamin D, naphtyl, isobutyl, benzyl, indol, tryptophane, and phenyl, which confer stability and efficient cellular uptake in the absence of any transfection reagent or formulation. Immune response assays testing for IFN-induced proteins indicate sdRNAs produce a reduced immunostimulatory profile as compared other RNAi agents. See, for example, Byrne et al., December 2013, J. Ocular Pharmacology and Therapeutics, 29(10): 855-864.

Cell-Based Immunotherapeutics

In general, cells are obtained from subjects with proliferative disease such as cancer, or an infectious disease such as viral infection. The obtained cells are treated directly as obtained or may be expanded in cell culture prior to treatment with oligonucleotides. The cells may also be genetically modified to express receptors that recognize specific antigens expressed on the tumor cell surface (CAR) or intracellular tumor antigens presented on MHC class I (TCR).

Oligonucleotide Agents

Antisense Oligonucleotides

Small interfering RNA (siRNA), sometimes known as short interfering RNA or silencing RNA, is a double stranded RNA molecule, generally 19-25 base pairs in length. siRNA is used in RNA interference (RNAi), where it interferes with expression of specific genes with complementary nucleotide sequences.

Double stranded DNA (dsRNA) can be generally used to define any molecule comprising a pair of complementary strands of RNA, generally a sense (passenger) and antisense (guide) strands, and may include single-stranded overhang regions. The term dsRNA, contrasted with siRNA, generally refers to a precursor molecule that includes the sequence of an siRNA molecule which is released from the larger dsRNA molecule by the action of cleavage enzyme systems, including Dicer.

sdRNA (self-deliverable) are a new class of covalently modified RNAi compounds that do not require a delivery vehicle to enter cells and have improved pharmacology compared to traditional siRNAs. “Self-deliverable RNA” or sdRNA is a hydrophobically modified RNA interfering-antisense hybrid, demonstrated to be highly efficacious in vitro in primary cells and in vivo upon local administration. Robust uptake and/or silencing without toxicity has been demonstrated in several tissues including dermal, muscle, tumors, alveolar macrophages, spinal cord, and retina cells and tissues. In dermal layer and retina, intradermal and intra-vitreal injection of sdRNA at mg doses induced potent and long lasting silencing.

While sdRNA is a superior functional genomics tool, enabling RNAi in primary cells and in vivo, it has a relatively low hit rate as compared to conventional siRNAs. While the need to screen large number of sequences per gene is not a limiting factor for therapeutic applications, it severely limits the applicability of sdRNA technology to functional genomics, where cost effective compound selection against thousands of genes is required. To optimize sdRNA structure, chemistry, targeting position, sequence preferences, and the like, a proprietary algorithm has been developed and utilized for sdRNA potency prediction. Availability of sdRNA reagents that are active in all cell types ex vivo and in vivo enables functional genomics and target stratification/validation studies.

Proprietary Algorithm

SdRNA sequences were selected based on a proprietory selection algorithm, designed on the basis of a functional screen of over 500 sdRNA sequences in the luciferase reporter assay of HeLa cells. Regression analysis of was used to establish a correlation between the frequency of occurrence of specific nucleotide and modification at any specific position in sdRNA duplex and its functionality in gene suppression assay. This algorithm allows prediction of functional sdRNA sequences, defined as having over 70% knockdown at 1 μM concentration, with a probability over 40%.

Table 1 shows predictive gene targets identified using the proprietary algorithm and useful in the cellular immunotherapeutic compositions and methods described herein.

Delivery of RNAi Agents

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; Applic BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; ation of RNAi technology to functional genomics studies in prim BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; ary cells and in vivo is limited by requirements to formulate siRNAs into lipids or use of other cell delivery techniques. To circumvent delivery problems, the self-deliverable RNAi technology provides a method of directly transfecting cells with the RNAi agent, without the need for additional formulations or techniques. The ability to transfect hard-to-transfect cell lines, high in vivo activity, and simplicity of use, are characteristics of the compositions and methods that present significant functional advantages over traditional siRNA-based techniques. The sdRNAi technology allows direct delivery of chemically synthesized compounds to a wide range of primary cells and tissues, both ex-vivo and in vivo.

To enable BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; self-delivery, traditional siRNA molecules require a substantial reduction in size and the introduction of extensive chemical modifications which are not well tolerated by RNAi machinery, resulting in extremely low probability of finding active molecules (low hit rate). In contrast, the sdRNA technology allows efficient RNAi delivery to primary cells and tissues in vitro and in vivo, with demonstrated silencing efficiency in humans.

The general structure of sdRNA molecules is shown in FIG. 1. sdRNA are formed as hydrophobically-modified siRNA-antisense oligonucleotide hybrid structures, and are disclosed, for example in Byrne et al., December 2013, J. Ocular Pharmacology and Therapeutics, 29(10): 855-864.

Oligonucleotide Modifications: 2′-O-Methyl, 2′-O-Fluro, Phosphorothioate

The oligonucleotide agents preferably comprise one or more modification to increase stability and/or effectiveness of the therapeutic agent, and to effect efficient delivery of the oligonucleotide to the cells or tissue to be treated. Such modifications include at least one

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; 2′-O-methyl modification, at least one 2′-O-Fluro modification, and at least one diphosphorothioate modification. Additionally, the oligonucleotide is modified to include one or more hydrophobic modification selected from sterol, cholesterol, vitamin D, naphtyl, isobutyl, benzyl, indol, tryptophane, and phenyl. The hydrophobic modification is preferably a sterol.

Delivery of Oligonucleotide Agents to Cells

The oligonucleotides may be delivered to the cells in combination with a transmembrane delivery system, preferably comprising lipids, viral vectors, and the like. Most preferably, the oligonucleotide agent is a self-delivery RNAi agent, that does not require any delivery agents.

Combination Therapy

Most preferred for this invention, e.g. particular combinations of elements and/or alternatives for specific needs. This objective is accomplished by determining the appropriate genes to be targeted by the oligonucleotide in order to silence immune suppressor genes and using the proprietary algorithm to select the most appropriate target sequence.

It is preferred that the immunotherapeutic cell be modified to include multiple oligonucleotide agents targeting a variety of genes involved in immune suppression and appropriate for the selected target disease and genes. For example, a preferred immunotherapeutic cell is a T-Cell modified to knock-down both CTLA-4 and PD-1

Additional combinations of oligonucleotides to related genes involved in immune suppression include varied combinations of the selected target sequences of Table 1.

BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; Preferred BTLA (B and T-lymphocyte attenuator), KIR (killer immunoglobulin-like receptors), B7-H3 and B7-H4 receptors and TGFbeta type 2 receptor; therapeutic combinations include cells engineered to knock down gene expression of the following target genes:

-   -   a) CTLA4 and PD1     -   b) STAT3 and p38     -   c) PD1 and BaxPD1, CTLA4, Lag-1, ILM-3, and TP53     -   d) PD1 and Casp8     -   e) PD1 and IL10R

The therapeutic compositions described herein are useful to treat a subject suffering from a proliferation disorder or infectious disease. In particular, the immunotherapeutic composition is useful to treat disease characterized by suppression of the subjects immune mechanisms. The sdRNA agents described herein are specifically designed to target genes involved in diseases-associated immune suppression pathways.

Methods of treating a subject comprise administering to a subject in need thereof, an immunogenic composition comprising an sdRNAi agent capable of inhibiting expression of genes involved in immune suppression mechanisms, for example, any of the genes listed in Table 1 or otherwise described herein.

EXAMPLES

The embodiments of the invention described above are intended to be merely exemplary; numerous variations and modifications will be apparent to those skilled in the art. All such variations and modifications are intended to be within the scope of the present invention as defined in any appended claims.

Example 1 Self-Deliverable RNAi Immunotherapeutic Agents

Immunotherapeutic agents described herein were produced by treating cells with particular sdRNA agents designed to target and knock down specific genes involved in immune suppression mechanisms. In particular, the following cells and cell lines have been successfully treated with sdRNA and were shown to knock down at least 70% of targeted gene expression in the specified human cells.

These studies demonstrated utility of these immunogenic agents to suppress expression of target genes in cells normally very resistant to transfection, and suggests the agents are capable of reducing expression of target cells in any cell type.

TABLE 2 Cell Type Target Gene sdRNA target sequence % Knock Down Primary human T-cells TP53 (P53) GAGTAGGACATACCAGCTTA (SEQ ID NO: 1001) >70% 2 uM Primary human T-cells MAP4K4 AGAGTTCTGTGGAAGTCTA (SEQ ID NO: 1002) >70% 2 uM Jurkat T-Iymphoma cells MAP4K4 AGAGTTCTGTGGAAGTCTA (SEQ ID NO: 1003) 100% 1 uM 72h NK-92 cells MAP4K4 AGAGTTCTGTGGAAGTCTA (SEQ ID NO: 1004)  80% 2 uM 72h NK-92 cells PPIB ACAGCAAATTCCATCGTGT (SEQ ID NO: 1005) >75% 2 uM 72h NK-92 cells GADPH CTGGTAAAGTGGATATTGTT (SEQ ID NO: 1006) >90% 2 uM 72h HeLa Cells MAP4K4 AGAGTTCTGTGGAAGTCTA (SEQ ID NO: 1007) >80% 2 uM 72h

Example 2 Oligonucleotide Sequences for Inhibiting Expression of Target Genes

A number of human genes were selected as candidate target genes due to involvement in immune suppression mechanisms, including the following genes shown in Table 3:

TABLE 3 BAX (NM_004324) BAK1 (NM_001188) CASP8 (NM_001228) ADORA2A (NM_000675) CTLA4 (NM_005214) LAG3 (NM002286) PDCD1 (NM_NM005018) TGFBR1 (NM-004612) HAVCR2 (NM_032782) CCL17 (NM_002987) CCL22 (NM_002990) DLL2 (NM_005618) FASLG (NM_000639) CD274 (NM_001267706) IDO1 (NM_002164) IL10RA (NM_001558) JAG1 (NM_000214) JAG2 (NM_002226) MAPK14 (NM_001315) SOCS1 (NM_003745) STAT3 (NM_003150) TNFA1P3 (NM_006290) TNFSF4 (NM_003326) TYRO2 (NM_006293) TP53 (NM_000546)

Each of the genes listed above was analyzed using a proprietary algorithm to identify preferred sdRNA targeting sequences and target regions for each gene for prevention of immunosuppression of antigen-presenting cells and T-cells. Results are shown in Table 1.

Example 3 Knock-Down of Target Gene (GAPDH) by sdRNA in HeLa Cells

HeLa cells (ATCC CRM-CCL-2) were subcultured 24 hours before transfection and kept log phase. The efficacy of several GAPDH sdRNAs was tested by qRT-PCR, including G13 sdRNA listed in the Table 1.

Solutions of GAPDH, MAP4K4 (positive control) and NTC (non-targeting control) sdRNA with twice the required concentration were prepared in serum-free EMEM medium, by diluting 100 μM oligonucleotides to 0.2-4 μM.

The total volume of medium for each oligo concentration point was calculated as [50 μl/well]×[number of replicates for each serum point]. Oligonucleotides were dispensed into a 96 well plate at 50 μl/well.

Cells were collected for transfection by trypsinization in a 50 ml tube, washed twice with medium containing 10% FBS without antibiotics, spun down at 200×g for 5 minutes at room temperature and resuspended in EMEM medium containing twice the required amount of FBS for the experiment (6%) and without antibiotics. The concentration of the cells was adjusted to 120,000/ml to yield a final concentration of 6,000 cells/50 μl/well. The cells were dispensed at 50 μl/well into the 96-well plate with pre-diluted oligos and placed in the incubator for 48 hours.

Gene Expression Analysis in HeLa Cells Using qRT-PCR

RNA was isolated from transfected HeLa cells using the PureLink™ Pro96 total RNA purification Kit (Ambion, Cat. No. 12173-011A), with Quanta qScript XLT One-Step RT-qPCR ToughMix, ROX (VWR, 89236672). The isolated RNA was analyzed for gene expression using the Human MAP4K4-FAM (Taqman Hs0377405 ml) and Human GAPDH-VIC (Applied Biosystems, Cat. No. 4326317E) gene expression assays.

The incubated plate was spun down and washed once with 100 μl/well PBS and lysed with 60 μl/well buffer provided in the kit. RNA isolation was conducted according to the manufacturer's instructions, and the RNA was eluted with 100 μl RNase-free water, and used undiluted for one-step qRT-PCR.

Dilutions of non-transfected (NT) cells of 1:5 and 1:25 were prepared for the standard curve using RNase-free water. qRT-PCR was performed by dispensing 9 μl/well into a low profile PCR plate and adding 1 μl RNA/well from the earlier prepared RNA samples. After brief centrifugation, the samples were placed in the real-time cycler and amplified using the settings recommended by the manufacturer.

GAPDH gene expression was measured by qPCR, normalized to MAP4K4 and plotted as percent of expression in the presence of non-targeting sdRNA. The results were compared to the normalized according to the standard curve. As shown in FIG. 2, several sdRNA agents targeting GAPDH or MAP4K4 significantly reduced their mRNA levels leading to more than 80-90% knock-down with 1 μM sdRNA. (See FIG. 2).

Example 4 Silencing of Multiple Targets by sdRNA in NK-92 Cells

NK-92 cells were obtained from Conqwest and subjected to one-step RT-PCR analysis without RNA purification using the FastLane Cell Multiplex Kit (Qiagen, Cat. No. 216513). For transfection, NK-92 cells were collected by centrifugation and diluted with RPMI medium containing 4% FBS and IL2 1000 U/ml and adjusted to1,000,000 cells/ml.

Multiple sdRNA agents targeting MAP4K4, PPIB or GADPH were diluted separately in serum-free RPMI medium to 4 μM and individually aliquoted at 50 μl/well into a 96-well plate. The prepared cells were then added at 50 μl cells/well to the wells with either MAP4K4, PPIB or GAPDH sdRNAs. Cells were incubated for 24, 48, or 72 hours.

At the specified timepoints, the plated transfected cells were washed once with 100 μl/well PBS and once with FCW buffer. After removal of supernatant, cell processing mix of 23.5 μl FCPL and 1.5 μl gDNA wipeout solution was added to each well and incubated for five minutes at room temperature. Lysates were then transferred to PCR strips and heated at 75° C. for five minutes.

To setup qRT-PCR, the lysates were mixed with QuantiTect reagents from the FastLane Cell Multiplex Kit and with primer probe mix for MAP4K4-FAM/GAPDH-VIC or PPIB-FAM/GAPDH-VIC. The following Taqman gene expression assays were used: human MAP4K4-FAM (Taqman, Hs00377405_m1), human PPIB-FAM (Taqman, Hs00168719_m1) and human GAPDH-VIC (Applied Biosystems, cat. No 4326317E).

A volume of 9 μl/well of each reaction mix was dispensed into a low profile PCR plate. One μl lysate per well was added from the previously prepared lysates. The samples were amplified using the settings recommended by the manufacturer.

Results shown in FIG. 3 demonstrate significant silencing of each of the multiple targets, MAP4K4, PPIB, and GADPH by sdRNA agents transfected into NK-92 cells, including greater than 75% inhibition of expression of each target within 24 to 72 hours of incubation.

Example 5 Silencing of TP53 and MAP4K4 by sdRNA in Human Primary T-Cells

Primary human T-cells were obtained from AllCells (CA) and cultured in complete RPMI medium containing 1000 IU/ml IL2. Cells were activated with anti-CD3/CD28 Dynabeads (Gibco, 11131) according to the manufacturer's instructions for at least 4 days prior to the transfection. Cells were collected by brief vortexing to dislodge the beads from cells and separating them using the designated magnet.

sdRNA agents targeting TP53 or MAP4K4 were prepared by separately diluting the sdRNAs to 0.2-4 μM in serum-free RPMI per sample (well) and individually aliquoted at 100 μl/well of 96-well plate. Cells were prepared in RPMI medium containing 4% FBS and IL2 2000 U/ml at 1,000,000 cells/ml and seeded at 100 μl/well into the 96-well plate with pre-diluted sdRNAs.

At the end of the transfection incubation period, the plated transfected cells were washed once with 100 μl/well PBS and processed with FastLane Cell Multiplex Kit reagents essentially as described for the Example 4 and according to the manufacturer's instructions. Taqman gene expression assays were used in the following combinations: human MAP4K4-FAM/GAPDH-VIC or human TP53-FAM (Taqman, Hs01034249_m1)/GAPDH-VIC. A volume of 18 μl/well of each reaction mix was combined with 2 μl lysates per well from the previously prepared lysates. The samples were amplified as before (see Example 4).

Results shown in FIG. 4 demonstrate significant silencing of both MAP4K4 and TP53 by sdRNA agents transfected into T-cells, reaching 70-80% inhibition of gene expression with 1-2 μM sdRNA.

Example 6 Immunotherapeutic Combination of sdRNAs for Treating Melanoma

Melanomas utilize at least two particular pathways to suppress immune function of T-cells, and each involves both PD1 and CTLA4. Melanoma tumors expressing the PD1 ligand, PD1L, can be targeted with T-cells pretreated ex-vivo with sd-RNAi agents specifically designed to target PD1 and interfere with PD1 expression. PD1 is also known as PDCD1, and particular targeting sequences and gene regions identified and predicted to be particularly functional in sdRNA mediated suppression, are shown in Table 1 for PDCD1 (NM_005018) and for CTLA4 (NM005214).

Treatment of melanoma tumors can be effected by providing to melanoma cells T-cells, such as tumor-infiltrating lymphocytes, pretreated ex-vivo with a combination of sdRNAs targeting PD1/PDCD1 and CTLA4, for example, targeting one or more of the twenty target sequences listed for PD1/PDCD1 and/or CTLA4. A combination of sdRNAs targeting PD1/PDCD1 and FASLG (NM_000639) and/or CTLA4, can increase T-cell toxicity in tumors expressing both PD1L and FAS.

In addition to and in combination with anti-CTLA-4 and anti-PD1 sdRNAs, T-cells used for the immunotherapy of melanoma can also be treated with sdRNA targeting other genes implicated in immunosuppression by the tumor. These receptors include, but are not limited to TGF-beta type 1 and 2 receptors, BTLA (binder of herpes virus entry indicator (HVEM) expressed on melanoma cells), and receptors of integrins expressed by myeloid derived suppressor cells (MDSC), such as CD11b, CD18, and CD29.

For tumors whose profile of expressed suppressive proteins is unknown, any combination of sdRNAs targeting PD1/PDCD1 and any one of know suppressing receptors may be helpful to reduce immune suppression and increase therapeutic efficacy.

Example 7 Combination of sdRNAs for Mitigating Immune Cell Suppression

T-cell or dendritic cell suppression may be modulated by various cytokines, such as IL10 and/or TGF beta. Suppressing corresponding receptors in T-cells and dendritic cells may be beneficial for their activity. For example, providing a combination of anti-PD1 with anti-IL10R sdRNAs is exptected to mitigate cytokine induced suppression of T-cells and dendritic cells, as compared with anti-PD1 alone.

Example 8 Combination of sdRNAs for Mitigating Immune Cell Suppression

When the mechanism of tumor suppression of immune cells may be not known, use of sdRNA agents to suppress genes involved in apoptosis (programmed cell death), such as p53, Casp8 or other gene activating apoptosis may be beneficial to increase immune cell activity. Combination of an anti-receptor sdRNAs with sdRNAs against pro-apoptotic genes can additionally reduce death of immune cells and thus increase their activity. For example, combination of anti-PD1 with anti-p53 sdRNAs may additionally protect T-cells from suppression by blocking activation of apoptosis.

Example 9 Silencing of CTLA-4 and PDCD1 by sdRNA in Human Primary T-Cells

Primary human T-cells were cultured and activated essentially as described in Example 5. sdRNA agents targeting PDCD1 and CTLA-4 were prepared by separately diluting the sdRNAs to 0.4-4 μM in serum-free RPMI per sample (well) and aliquoted at 100 μl/well of 96-well plate. Cells were prepared in RPMI medium containing 4% FBS and IL2 2000 U/ml at 1,000,000 cells/ml and seeded at 100 μl/well into the 96-well plate with pre-diluted sdRNAs.

72 h later, the transfected cells were washed once with 100 μl/well PBS and processed with FastLane Cell Multiplex Kit reagents essentially as described for the Example 4 and according to the manufacturer's instructions. Taqman gene expression assays were used in the following combinations: human PDCD1-FAM (Taqman, Hs01550088_m1)/GAPDH-VIC or human CTLA4-FAM (Taqman, Hs03044418_m1)/GAPDH-VIC. A volume of 18 μl/well of each reaction mix was combined with 2 μl lysates per well from the previously prepared lysates. The samples were amplified as before (see Example 4).

Results shown in FIG. 5 demonstrate significant silencing of PDCD1 and CTLA-4 by using combined sdRNA agents delivered to T-cells, obtaining greater than 60-70% inhibition of gene expression with 2 μM sdRNA.

Example 10 Reduction of CTLA-4 and PDCD1 Surface Expression by sdRNA in Human Primary T-Cells

Primary human T-cells were cultured and activated essentially as described in Example 5.

sdRNA agents targeting CTLA-4 or PD1 were separately diluted to 5 μM in serum-free RPMI per sample (well) and aliquoted at 250 μl/well to 24-well plates. Cells mixed with magnetic beads were collected and adjusted to 500,000 cells in 250 μl RPMI medium containing 4% FBS and IL2 2000 IU/ml. Cells were seeded at 250 μl/well to the prepared plate containing pre-diluted sdRNAs. 24 hours later FBS was added to the cells to obtain 10% final concentration.

After 72 hours of incubation, the transfected cells were collected, separated from the activation beads using the magnet, as described in Example 5. Cells were washed with PBS, spun down and resuspended in blocking buffer (PBS with 3% BSA) at 200,000 cells/50 μl/sample.

Antibody dilutions were prepared in the blocking buffer. The antibodies were mixed in two combinations: anti-PD1/anti-CD3 (1:100 dilutions for both antibodies) and anti-CTLA4/anti-CD3 (10 μl/106 cells for anti-CTLA4; 1:100 for CD3). The following antibodies were used: rabbit monoclonal [SP7] to CD3 (Abcam, ab16669); mouse monoclonal [BNI3] to CTLA4 (Abcam, ab33320) and mouse monoclonal [NAT105] to PD1 (Abcam, ab52587). Cells were mixed with the diluted antibodies and incubated 30 minutes on ice. Cells were then washed twice with PBS containing 0.2% Tween-20 and 0.1% sodium azide.

Secondary antibodies were diluted in blocking buffer and mixed together resulting in a final dilution 1:500 for anti-mouse Cy5 (Abcam, ab97037) and 1:2000 for anti-rabbit Alexa-488 (Abcam, ab150077). Cells were mixed with the diluted antibodies at 1:1 ratio and incubated 30 minutes on ice. Cells were washed as before, and diluted in 500 μl PBS per tube. The data was acquired immediately on the Attune Acoustic Focusing Cytometer (Applied Biosystems).

As shown in FIG. 6, sdRNA efficiently reduced surface expression of CTLA-4 and PD1 in activated Human Primary T cells.

Example 11 MAP4K4 sdRNA Delivery into CD3- and CD19-Positive Subsets of Human Peripheral Blood Mononuclear Cells (PBMCs)

PBMCs were cultured in complete RPMI supplemented with 1.5% PHA solution and 500 U/ml IL2. For transfection, PBMCs were collected by centrifugation and diluted with RPMI medium containing 4% FBS and IL2 1000 U/ml and seeded to 24-well plate at 500,000 cells/well.

MAP4K4 sdRNA labeled with cy3 was added to the cells at 0.1 μM final concentration. After 72 hours of incubation, the transfected cells were collected, washed with PBS, spun down and diluted in blocking buffer (PBS with 3% BSA) at 200,000 cells/50 μl/sample.

Antibody dilutions were prepared in the blocking buffer as following: 1:100 final dilution anti-CD3 (Abcam, ab16669) and anti-CD19 at 10 μl/1,000,000 cells (Abcam, ab31947). Cells were mixed with the diluted antibodies and incubated 30 min on ice. Cells were then washed twice with PBS containing 0.2% Tween-20 and 0.1% sodium azide.

Secondary antibodies were diluted in the blocking buffer in a final dilution 1:500 for anti-mouse Cy5 (Abcam, ab97037) and 1:2000 for anti-rabbit Alexa-488 (Abcam, ab150077). Cells were mixed with the diluted antibodies at 1:1 ratio and incubated 30 min on ice. Cells were washed as before, and diluted in 500 μl PBS per tube. The data was acquired immediately on the Attune Acoustic Focusing Cytometer (Applied Biosystems).

FIG. 7 shows efficient transfection over 97% of CD3-positive (t cells) and over 98% CD19-positive (B-cells) subsets in Human Peripheral Blood Mononuclear Cells (PBMCs).

TABLE 1 Targeting sequences and gene regions of genes targeted with sdRNAs to prevent immunosuppression of antigen-presenting cells and T-cells. SEQ ID SEQ ID Oligo_count Oligo_ID targeting sequence NO: Gene_region NO: Accession: NM_004324 HUGO gene symbol: BAX 1 BAX_NM_004324_ GAATTGCTCAAGTTCATTGA 1 CCTCCACTGCCTCTGGAATTGCTCAAG 21 human_835 TTCATTGATGACCCTCTG 2 BAX_NM_004324_ TTCATCCAGGATCGAGCAGG 2 CTTTTGCTTCAGGGTTTCATCCAGGAT 22 human_ 57 CGAGCAGGGCGAATGGGG 3 BAX_NM_004324_ ATCATCAGATGTGGTCTATA 3 TCTCCCCATCTTCAGATCATCAGATGT 23 human_684 GGTCTATAATGCGTTTTC 4 BAX_NM_004324_ TACTTTGCCAGCAAACTGGT 4 GTTGTCGCCCTTTTCTACTTTGCCAGCA 24 human_412 AACTGGTGCTCAAGGCC 5 BAX_NM_004324_ GGTTGGGTGAGACTCCTCAA 5 ATCCAAGACCAGGGTGGTTGGGTGAG 25 human_538 ACTCCTCAAGCCTCCTCAC 6 BAX_NM_004324_ CTACTTTGCCAGCAAACTGG 6 GGTTGTCGCCCTTTTCTACTTTGCCAGC 26 human_411 AAACTGGTGCTCAAGGC 7 BAX_NM_004324_ GCGTTTTCCTTACGTGTCTG 7 GATGTGGTCTATAATGCGTTTTCCTTA 27 human_706 CGTGTCTGATCAATCCCC 8 BAX_NM_004324_ TACGTGTCTGATCAATCCCC 8 ATAATGCGTTTTCCTTACGTGTCTGATC 28 human_7 16 AATCCCCGATTCATCTA 9 BAX_NM_004324_ TCAGGGTTTCATCCAGGATC 9 AGGGGCCCTTTTGCTTCAGGGTTTCAT 29 human_ 50 CCAGGATCGAGCAGGGCG 10 BAX_NM_004324_ TGACGGCAACTTCAACTGGG 10 AGCTGACATGTTTTCTGACGGCAACTT 30 human_ 72 CAACTGGGGCCGGGTTGT 11 BAX_NM_004324_ CAGCTGACATGTTTTCTGAC 11 TCTTTTTCCGAGTGGCAGCTGACATGT 31 human_356 TTTCTGACGGCAACTTCA 12 BAX_NM_004324_ AGCTGACATGTTTTCTGACG 12 CTTTTTCCGAGTGGCAGCTGACATGTT 32 human_357 TTCTGACGGCAACTTCAA 13 BAX_NM_004324_ CACTGTGACCTTGACTTGAT 13 AGTGACCCCTGACCTCACTGTGACCTT 33 human_776 GACTTGATTAGTGCCTTC 14 BAX_NM_004324_ TCCTTACGTGTCTGATCAAT 14 GTCTATAATGCGTTTTCCTTACGTGTCT 34 human_712 GATCAATCCCCGATTCA 15 BAX_NM_004324_ GATCAGAACCATCATGGGCT 15 CAAGGTGCCGGAACTGATCAGAACCA 35 human_465 TCATGGGCTGGACATTGGA 16 BAX_NM_004324_ CTTCTGGAGCAGGTCACAGT 16 TCTGGGACCCTGGGCCTTCTGGAGCA 36 human_642 GGTCACAGTGGTGCCCTCT 17 BAX_NM_004324_ TGAGCAGATCATGAAGACAG 17 GGGGCCCACCAGCTCTGAGCAGATCA 37 human_117 TGAAGACAGGGGCCCTTTT 18 BAX_NM_004324_ TATAATGCGTTTTCCTTACG 18 TCATCAGATGTGGTCTATAATGCGTTT 38 human_700 TCCTTACGTGTCTGATCA 19 BAX_NM_004324_ CCCATCTTCAGATCATCAGA 19 CAGTGGTGCCCTCTCCCCATCTTCAGA 39 human_673 TCATCAGATGTGGTCTAT 20 BAX_NM_004324_ AGGTGCCGGAACTGATCAGA 20 AGGCCCTGTGCACCAAGGTGCCGGAA 40 human_452 CTGATCAGAACCATCATGG Accession: NM_001188 HUGO gene symbol: BAK1 1 BAK1_NM_001188 TGGTTTGTTATATCAGGGAA 41 ACAGGGCTTAGGACTTGGTTTGTTA 61 _human_1813 TATCAGGGAAAAGGAGTAGG 2 BAK1_NM_001188 TGGTACGAAGATTCTTCAAA 42 TGTTGGGCCAGTTTGTGGTACGAAG 62 _human_911 ATTCTTCAAATCATGACTCC 3 BAK1_NM_001188 TTATATCAGGGAAAAGGAGT 43 TTAGGACTTGGTTTGTTATATCAGG 63 _human_1820 GAAAAGGAGTAGGGAGTTCA 4 BAK1_NM_001188 TCCCTTCCTCTCTCCTTATA 44 GTCCTCTCAGTTCTCTCCCTTCCTCTC 64 _human_1678 TCCTTATAGACACTTGCT 5 BAK1_NM_001188 TCAAATCATGACTCCCAAGG 45 TGGTACGAAGATTCTTCAAATCATG 65 _human_926 ACTCCCAAGGGTGCCCTTTG 6 BAK1_NM_001188 TGTTATATCAGGGAAAAGGA 46 GCTTAGGACTTGGTTTGTTATATCA 66 _human_1818 GGGAAAAGGAGTAGGGAGTT 7 BAK1_NM_001188 ACGAAGATTCTTCAAATCAT 47 GGGCCAGTTTGTGGTACGAAGATTC 67 _human_915 TTCAAATCATGACTCCCAAG 8 BAK1_NM_001188 GGTACGAAGATTCTTCAAAT 48 GTTGGGCCAGTTTGTGGTACGAAGA 68 _human_912 TTCTTCAAATCATGACTCCC 9 BAK1_NM_001188 GAAGTTCTTGATTCAGCCAA 49 GGGGGTCAGGGGGGAGAAGTTCTT 69 _human_2086 GATTCAGCCAAATGCAGGGAG 10 BAK1_NM_001188 CCTATGAGTACTTCACCAAG 50 CCACGGCAGAGAATGCCTATGAGTA 70 _human_620 CTTCACCAAGATTGCCACCA 11 BAK1_NM_001188 TATCAGGGAAAAGGAGTAGG 51 GGACTTGGTTTGTTATATCAGGGAA 71 _human_1823 AAGGAGTAGGGAGTTCATCT 12 BAK1_NM_001188 CTCTCCTTATAGACACTTGC 52 GTTCTCTCCCTTCCTCTCTCCTTATAG 72 _human_1687 ACACTTGCTCCCAACCCA 13 BAK1_NM_001188 ACTTGGTTTGTTATATCAGG 53 ACTACAGGGCTTAGGACTTGGTTTG 73 _human_1810 TTATATCAGGGAAAAGGAGT 14 BAK1_NM_001188 AAGATCAGCACCCTAAGAGA 54 ATTCAGCTATTCTGGAAGATCAGCA 74 _human_1399 CCCTAAGAGATGGGACTAGG 15 BAK1_NM_001188 GTTTGAGAGTGGCATCAATT 55 GATTGCCACCAGCCTGTTTGAGAGT 75 _human_654 GGCATCAATTGGGGCCGTGT 16 BAK1_NM_001188 GACTATCAACACCACTAGGA 56 TCTAAGTGGGAGAAGGACTATCAAC 76 _human_1875 ACCACTAGGAATCCCAGAGG 17 BAK1_NM_001188 AGCTTTAGCAAGTGTGCACT 57 CCTCAAGAGTACAGAAGCTTTAGCA 77 _human_1043 AGTGTGCACTCCAGCTTCGG 18 BAK1_NM_001188 TTCATCTGGAGGGTTCTAAG 58 AAAAGGAGTAGGGAGTTCATCTGG 78 _human_1846 AGGGTTCTAAGTGGGAGAAGG 19 BAK1_NM_001188 AAGTTCTTGATTCAGCCAAA 59 GGGGTCAGGGGGGAGAAGTTCTTG 79 _human_2087 ATTCAGCCAAATGCAGGGAGG 20 BAK1_NM_001188 GTTATATCAGGGAAAAGGAG 60 CTTAGGACTTGGTTTGTTATATCAG 80 _human_1819 GGAAAAGGAGTAGGGAGTTC Accession: NM_001228 HUGO gene symbol: CASP8 1 CASP8_NM_001228 TTAAATCATTAGGAATTAAG 121 TCTGCTTGGATTATTTTAAATCATTAG 141 _human_2821 GAATTAAGTTATCTTTAA 2 CASP8_NM_001228 GAATTAAGTTATCTTTAAAA 122 ATTTTAAATCATTAGGAATTAAGTTAT 142 _human_2833 CTTTAAAATTTAAGTATC 3 CASP8_NM_001228 AACTTTAATTCTCTTTCAAA 123 TGTTAATATTCTATTAACTTTAATTCT 143 _human_2392 CTTTCAAAGCTAAATTCC 4 CASP8_NM_001228 GACTGAAGTGAACTATGAAG 124 TATTCTCACCATCCTGACTGAAGTGA 144 _human_1683 ACTATGAAGTAAGCAACAA 5 CASP8_NM_001228 ATATTCTCCTGCCTTTTAAA 125 GGGAATATTGAGATTATATTCTCCTG 145 _human_281 CCTTTTAAAAAGATGGACT 6 CASP8_NM_001228 AGTTATCTTTAAAATTTAAG 126 AATCATTAGGAATTAAGTTATCTTTA 146 _human_2839 AAATTTAAGTATCTTTTTT 7 CASP8_NM_001228 TAGATTTTCTACTTTATTAA 127 TATTTACTAATTTTCTAGATTTTCTACT 147 _human_2164 TTATTAATTGTTTTGCA 8 CASP8_NM_001228 CTGTGCCCAAATCAACAAGA 128 CATCCTGAAAAGAGTCTGTGCCCAAA 148 _human_888 TCAACAAGAGCCTGCTGAA 9 CASP8_NM_001228 AGCTGGTGGCAATAAATACC 129 TTTGGGAATGTTTTTAGCTGGTGGCA 149 _human_2283 ATAAATACCAGACACGTAC 10 CASP8_NM_001228 TCCTACCGAAACCCTGCAGA 130 GTGAATAACTGTGTTTCCTACCGAAA 150 _human_1585 CCCTGCAGAGGGAACCTGG 11 CASP8_NM_001228 TATAAGAGCTAAAGTTAAAT 131 TGTTTTGCACTTTTTTATAAGAGCTAA 151 _human_2200 AGTTAAATAGGATATTAA 12 CASP8_NM_001228 CACTATGTTTATTTACTAAT 132 ACTATTTAGATATAACACTATGTTTAT 152 _human_2140 TTACTAATTTTCTAGATT 13 CASP8_NM_001228 ATTGTTATCTATCAACTATA 133 GGGCTTATGATTCAGATTGTTATCTA 153 _human_2350 TCAACTATAAGCCCACTGT 14 CASP8_NM_001228 TAACTGTGTTTCCTACCGAA 134 GATGGCCACTGTGAATAACTGTGTTT 154 _human_1575 CCTACCGAAACCCTGCAGA 15 CASP8_NM_001228 TAATTCTCTTTCAAAGCTAA 135 ATATTCTATTAACTTTAATTCTCTTTCA 155 _human_2397 AAGCTAAATTCCACACT 16 CASP8_NM_001228 TATATGCTTGGCTAACTATA 136 TGCTTTTATGATATATATATGCTTGGC 156 _human_2726 TAACTATATTTGCTTTTT 17 CASP8_NM_001228 CTCTGCTTGGATTATTTTAA 137 CATTTGCTCTTTCATCTCTGCTTGGAT 157 _human_2805 TATTTTAAATCATTAGGA 18 CASP8_NM_001228 ATGCTTGGCTAACTATATTT 138 TTTTATGATATATATATGCTTGGCTAA 158 _human_2729 CTATATTTGCTTTTTGCT 19 CASP8_NM_001228 ATAAGAGCTAAAGTTAAATA 139 GTTTTGCACTTTTTTATAAGAGCTAAA 159 _human_2201 GTTAAATAGGATATTAAC 20 CASP8_NM_001228 ATCTTTAAAATTTAAGTATC 140 ATTAGGAATTAAGTTATCTTTAAAATT 160 _human_2843 TAAGTATCTTTTTTCAAA Accession: NM_000675 HUGO gene symbol: ADORA2A 1 ADORA2A_NM_00067 TAACTGCCTTTCCTTCTAAA 161 GTGAGAGGCCTTGTCTAACTGCC 181 5_human_2482 TTTCCTTCTAAAGGGAATGTTT 2 ADORA2A_NM_00067 TTCCTTCTAAAGGGAATGTT 162 CTTGTCTAACTGCCTTTCCTTCTAA 182 5_human_2491 AGGGAATGTTTTTTTCTGAG 3 ADORA2A_NM_00067 GCCTTTCCTTCTAAAGGGAA 163 AGGCCTTGTCTAACTGCCTTTCCT 183 5_human_2487 TCTAAAGGGAATGTTTTTTTC 4 ADORA2A_NM_00067 TTTTCTGAGATAAAATAAAA 164 CTAAAGGGAATGTTTTTTTCTGAG 184 5_human_2512 ATAAAATAAAAACGAGCCACA 5 ADORA2A_NM_00067 CATCTCTTGGAGTGACAAAG 165 TCTCAGTCCCAGGGCCATCTCTTG 185 5_human_2330 GAGTGACAAAGCTGGGATCAA 6 ADORA2A_NM_00067 CATGGTGTACTTCAACTTCT 166 GGTCCCCATGAACTACATGGTGT 186 5_human_987 ACTTCAACTTCTTTGCCTGTGT 7 ADORA2A_NM_00067 CTAACTGCCTTTCCTTCTAA 167 AGTGAGAGGCCTTGTCTAACTGC 187 5_human_2481 CTTTCCTTCTAAAGGGAATGTT 8 ADORA2A_NM_00067 CTGATGATTCATGGAGTTTG 168 TGGAGCAGGAGTGTCCTGATGAT 188 5_human_1695 TCATGGAGTTTGCCCCTTCCTA 9 ADORA2A_NM_00067 CTCAGAGTCCTCTGTGAAAA 169 CCTGGTTTCAGGAGACTCAGAGT 189 5_human_264 CCTCTGTGAAAAAGCCCTTGGA 10 ADORA2A_NM_00067 AACGAGCCACATCGTGTTTT 170 CTGAGATAAAATAAAAACGAGCC 190 5_human_2531 ACATCGTGTTTTAAGCTTGTCC 11 ADORA2A_NM_00067 TCCTTCTAAAGGGAATGTTT 171 TTGTCTAACTGCCTTTCCTTCTAAA 191 5_human_2492 GGGAATGTTTTTTTCTGAGA 12 ADORA2A_NM_00067 CATGAACTACATGGTGTACT 172 TGAGGATGTGGTCCCCATGAACT 192 5_human_978 ACATGGTGTACTTCAACTTCTT 13 ADORA2A_NM_00067 AACTGCCTTTCCTTCTAAAG 173 TGAGAGGCCTTGTCTAACTGCCTT 193 5_human_2483 TCCTTCTAAAGGGAATGTTTT 14 ADORA2A_NM_00067 CAGATGTTTCATGCTGTGAG 174 TGGGTTCTGAGGAAGCAGATGTT 194 5_human_1894 TCATGCTGTGAGGCCTTGCACC 15 ADORA2A_NM_00067 CCCATGAACTACATGGTGTA 175 TTTGAGGATGTGGTCCCCATGAA 195 5_human_976 CTACATGGTGTACTTCAACTTC 16 ADORA2A_NM_00067 AGGCAGCAAGAACCTTTCAA 176 CGCAGCCACGTCCTGAGGCAGCA 196 5_human_1384 AGAACCTTTCAAGGCAGCTGGC 17 ADORA2A_NM_00067 GTCCTGATGATTCATGGAGT 177 GGATGGAGCAGGAGTGTCCTGAT 197 5_human_1692 GATTCATGGAGTTTGCCCCTTC 18 ADORA2A_NM_00067 GTACTTCAACTTCTTTGCCT 178 CATGAACTACATGGTGTACTTCAA 198 5_human_993 CTTCTTTGCCTGTGTGCTGGT 19 ADORA2A_NM_00067 TGTAAGTGTGAGGAAACCCT 179 TTTTTCCAGGAAAAATGTAAGTGT 199 5_human_2167 GAGGAAACCCTTTTTATTTTA 20 ADORA2A_NM_00067 CCTACTTTGGACTGAGAGAA 180 TGAGGGCAGCCGGTTCCTACTTT 200 5_human_1815 GGACTGAGAGAAGGGAGCCCCA Accession: NM_005214 HUGO gene symbol: CTLA4 1 CTLA4_NM_005214_ TGATTCTGTGTGGGTTCAAA 201 TCTATATAAAGTCCTTGATTCTGT 221 human_61 GTGGGTTCAAACACATTTCAA 2 CTLA4_NM_005214_ TTATTTGTTTGTGCATTTGG 202 GCTATCCAGCTATTTTTATTTGTTT 222 human_909 GTGCATTTGGGGGGAATTCA 3 CTLA4_NM_005214_ TGATTACATCAAGGCTTCAA 203 TCTTAAACAAATGTATGATTACAT 223 human_1265 CAAGGCTTCAAAAATACTCAC 4 CTLA4_NM_005214_ GATGTGGGTCAAGGAATTAA 204 GGGATGCAGCATTATGATGTGGG 224 human_1094 TCAAGGAATTAAGTTAGGGAAT 5 CTLA4_NM_005214_ CCTTTTATTTCTTAAACAAA 205 AAGTTAAATTTTATGCCTTTTATTT 225 human_1241 CTTAAACAAATGTATGATTA 6 CTLA4_NM_005214_ GATTACATCAAGGCTTCAAA 206 CTTAAACAAATGTATGATTACATC 226 human_1266 AAGGCTTCAAAAATACTCACA 7 CTLA4_NM_005214_ TCTGTGTGGGTTCAAACACA 207 TATAAAGTCCTTGATTCTGTGTGG 227 human_65 GTTCAAACACATTTCAAAGCT 8 CTLA4_NM_005214_ TTGATAGTATTGTGCATAGA 208 TATATATATTTTAATTTGATAGTAT 228 human_1405 TGTGCATAGAGCCACGTATG 9 CTLA4_NM_005214_ TGCCTTTTATTTCTTAAACA 209 TCAAGTTAAATTTTATGCCTTTTAT 229 human_1239 TTCTTAAACAAATGTATGAT 10 CTLA4_NM_005214_ TCCATGAAAATGCAACAACA 210 TTTAACTCAATATTTTCCATGAAA 230 human_1912 ATGCAACAACATGTATAATAT 11 CTLA4_NM_005214_ TTATTTCTTAAACAAATGTA 211 TAAATTTTATGCCTTTTATTTCTTA 231 human_1245 AACAAATGTATGATTACATC 12 CTLA4_NM_005214_ TTAATGGTTTGAATATAAAC 212 GTTTTTGTGTATTTGTTAATGGTTT 232 human_1449 GAATATAAACACTATATGGC 13 CTLA4_NM_005214_ ATGTGGGTCAAGGAATTAAG 213 GGATGCAGCATTATGATGTGGGT 233 human_1095 CAAGGAATTAAGTTAGGGAATG 14 CTLA4_NM_005214_ AGCCGAAATGATCTTTTCAA 214 GTATGAGACGTTTATAGCCGAAA 234 human_1208 TGATCTTTTCAAGTTAAATTTT 15 CTLA4_NM_005214_ GTTTGAATATAAACACTATA 215 GTGTATTTGTTAATGGTTTGAATA 235 human_1455 TAAACACTATATGGCAGTGTC 16 CTLA4_NM_005214_ TATGCCTTTTATTTCTTAAA 216 TTTCAAGTTAAATTTTATGCCTTTT 236 human_1237 ATTTCTTAAACAAATGTATG 17 CTLA4_NM_005214_ TTCCATGAAAATGCAACAAC 217 TTTTAACTCAATATTTTCCATGAA 237 human_1911 AATGCAACAACATGTATAATA 18 CTLA4_NM_005214_ CATCTCTCTTTAATATAAAG 218 CATTTGGGGGGAATTCATCTCTCT 238 human_937 TTAATATAAAGTTGGATGCGG 19 CTLA4_NM_005214_ GGAATTCATCTCTCTTTAAT 219 TTTGTGCATTTGGGGGGAATTCAT 239 human_931 CTCTCTTTAATATAAAGTTGG 20 CTLA4_NM_005214_ ATCTATATAAAGTCCTTGAT 220 TCTGGGATCAAAGCTATCTATATA 240 human_45 AAGTCCTTGATTCTGTGTGGG Accession: NM_002286 HUGO gene symbol: LAG3 1 LAG3_NM_002286 GACTTTACCCTTCGACTAGA 241 ACTGGAGACAATGGCGACTTTACC 261 _human_1292 CTTCGACTAGAGGATGTGAGC 2 LAG3_NM_002286 CAACGTCTCCATCATGTATA 242 CTACAGAGATGGCTTCAACGTCTC 262 _human_1096 CATCATGTATAACCTCACTGT 3 LAG3_NM_002286 GTCCTTTCTCTGCTCCTTTT 243 TTTCTCATCCTTGGTGTCCTTTCTCT 263 _human_1721 GCTCCTTTTGGTGACTGGA 4 LAG3_NM_002286 TCCAGTATCTGGACAAGAAC 244 GCTTTGTGAGGTGACTCCAGTATC 264 _human_1465 TGGACAAGAACGCTTTGTGTG 5 LAG3_NM_002286 ATTTTCTGCCTTAGAGCAAG 245 GTGGCGACCAAGACGATTTTCTGC 265 _human_1795 CTTAGAGCAAGGGATTCACCC 6 LAG3_NM_002286 TTTCACCTTTGGAGAAGACA 246 ACTGGAGCCTTTGGCTTTCACCTTT 266 _human_1760 GGAGAAGACAGTGGCGACCA 7 LAG3_NM_002286 CATTTTGAACTGCTCCTTCA 247 AGCCTCCGACTGGGTCATTTTGAA 267 _human_904 CTGCTCCTTCAGCCGCCCTGA 8 LAG3_NM_002286 TCATCACAGTGACTCCCAAA 248 CTGTCACATTGGCAATCATCACAGT 268 _human_1398 GACTCCCAAATCCTTTGGGT 9 LAG3_NM_002286 GCTTTCACCTTTGGAGAAGA 249 TGACTGGAGCCTTTGGCTTTCACCT 269 _human_1758 TTGGAGAAGACAGTGGCGAC 10 LAG3_NM_002286 CTTTGGCTTTCACCTTTGGA 250 TTTGGTGACTGGAGCCTTTGGCTTT 270 _human_1753 CACCTTTGGAGAAGACAGTG 11 LAG3_NM_002286 ATTTTGAACTGCTCCTTCAG 251 GCCTCCGACTGGGTCATTTTGAACT 271 _human_905 GCTCCTTCAGCCGCCCTGAC 12 LAG3_NM_002286 CACATTGGCAATCATCACAG 252 GCTCAATGCCACTGTCACATTGGC 272 _human_1387 AATCATCACAGTGACTCCCAA 13 LAG3_NM_002286 TTTCTGACCTCCTTTTGGAG 253 ACTGCCCCCTTTCCTTTTCTGACCTC 273 _human_301 CTTTTGGAGGGCTCAGCGC 14 LAG3_NM_002286 CGACTGGGTCATTTTGAACT 254 ATCTCTCAGAGCCTCCGACTGGGT 274 _human_895 CATTTTGAACTGCTCCTTCAG 15 LAG3_NM_002286 TACTTCACAGAGCTGTCTAG 255 CTTGGAGCAGCAGTGTACTTCACA 275 _human_1625 GAGCTGTCTAGCCCAGGTGCC 16 LAG3_NM_002286 ATTGGCAATCATCACAGTGA 256 CAATGCCACTGTCACATTGGCAATC 276 _human_1390 ATCACAGTGACTCCCAAATC 17 LAG3_NM_002286 CTGTTTCTCATCCTTGGTGT 257 GCAGGCCACCTCCTGCTGTTTCTCA 277 _human_1703 TCCTTGGTGTCCTTTCTCTG 18 LAG3_NM_002286 TTGTGAGGTGACTCCAGTAT 258 CCTGGGGAAGCTGCTTTGTGAGGT 278 _human_1453 GACTCCAGTATCTGGACAAGA 19 LAG3_NM_002286 TTTGGCTTTCACCTTTGGAG 259 TTGGTGACTGGAGCCTTTGGCTTTC 279 _human_1754 ACCTTTGGAGAAGACAGTGG 20 LAG3_NM_002286 TGGAGACAATGGCGACTTTA 260 TGACCTCCTGGTGACTGGAGACAA 280 _human_1279 TGGCGACTTTACCCTTCGACT Accession: NM_005018 HUGO gene symbol: PDCD1 1 PDCD1_NM_005018 TATTATATTATAATTATAAT 281 CCTTCCCTGTGGTTCTATTATATTAT 301 _human_2070 AATTATAATTAAATATGAG 2 PDCD1_NM_005018 TCTATTATATTATAATTATA 282 CCCCTTCCCTGTGGTTCTATTATATT 302 _human_2068 ATAATTATAATTAAATATG 3 PDCD1_NM_005018 CATTCCTGAAATTATTTAAA 283 GCTCTCCTTGGAACCCATTCCTGAA 303 _human_1854 ATTATTTAAAGGGGTTGGCC 4 PDCD1_NM_005018 CTATTATATTATAATTATAA 284 CCCTTCCCTGTGGTTCTATTATATT 304 _human_2069 ATAATTATAATTAAATATGA 5 PDCD1_NM_005018 AGTTTCAGGGAAGGTCAGAA 285 CTGCAGGCCTAGAGAAGTTTCAGG 305 _human_1491 GAAGGTCAGAAGAGCTCCTGG 6 PDCD1_NM_005018 TGTGGTTCTATTATATTATA 286 GGGATCCCCCTTCCCTGTGGTTCTA 306 _human_2062 TTATATTATAATTATAATTA 7 PDCD1_NM_005018 TGTGTTCTCTGTGGACTATG 287 CCCCTCAGCCGTGCCTGTGTTCTCT 307 _human_719 GTGGACTATGGGGAGCTGGA 8 PDCD1_NM_005018 CCCATTCCTGAAATTATTTA 288 GAGCTCTCCTTGGAACCCATTCCTG 308 _human_1852 AAATTATTTAAAGGGGTTGG 9 PDCD1_NM_005018 TGCCACCATTGTCTTTCCTA 289 TGAGCAGACGGAGTATGCCACCAT 309 _human_812 TGTCTTTCCTAGCGGAATGGG 10 PDCD1_NM_005018 AAGTTTCAGGGAAGGTCAGA 290 CCTGCAGGCCTAGAGAAGTTTCAG 310 _human_1490 GGAAGGTCAGAAGAGCTCCTG 11 PDCD1_NM_005018 CTGTGGTTCTATTATATTAT 291 AGGGATCCCCCTTCCCTGTGGTTCT 311 _human_2061 ATTATATTATAATTATAATT 12 PDCD1_NM_005018 TTCTATTATATTATAATTAT 292 CCCCCTTCCCTGTGGTTCTATTATA 312 _human_2067 TTATAATTATAATTAAATAT 13 PDCD1_NM_005018 TTTCAGGGAAGGTCAGAAGA 293 GCAGGCCTAGAGAAGTTTCAGGG 313 _human_1493 AAGGTCAGAAGAGCTCCTGGCT 14 PDCD1_NM_005018 CTTGGAACCCATTCCTGAAA 294 ACCCTGGGAGCTCTCCTTGGAACC 314 _human_1845 CATTCCTGAAATTATTTAAAG 15 PDCD1_NM_005018 TCCCTGTGGTTCTATTATAT 295 ACAAGGGATCCCCCTTCCCTGTGG 315 _human_2058 TTCTATTATATTATAATTATA 16 PDCD1_NM_005018 CCTGTGGTTCTATTATATTA 296 AAGGGATCCCCCTTCCCTGTGGTTC 316 _human_2060 TATTATATTATAATTATAAT 17 PDCD1_NM_005018 TGGAACCCATTCCTGAAATT 297 CCTGGGAGCTCTCCTTGGAACCCA 317 _human_1847 TTCCTGAAATTATTTAAAGGG 18 PDCD1_NM_005018 CCTTCCCTGTGGTTCTATTA 298 GGGACAAGGGATCCCCCTTCCCTG 318 _human_2055 TGGTTCTATTATATTATAATT 19 PDCD1_NM_005018 TTCCCTGTGGTTCTATTATA 299 GACAAGGGATCCCCCTTCCCTGTG 319 _human_2057 GTTCTATTATATTATAATTAT 20 PDCD1_NM_005018 CACAGGACTCATGTCTCAAT 300 CAGGCACAGCCCCACCACAGGACT 320 _human_1105 CATGTCTCAATGCCCACAGTG Accession: NM_004612 HUGO gene symbol: TGFBR1 1 TGFBR1_NM_004612 CCTGTTTATTACAACTTAAA 321 GTTAATAACATTCAACCTGTTTAT 341 _human_5263 TACAACTTAAAAGGAACTTCA 2 TGFBR1_NM_004612 CCATTGGTGGAATTCATGAA 322 TTGCTCGACGATGTTCCATTGGTG 342 _human_1323 GAATTCATGAAGATTACCAAC 3 TGFBR1_NM_004612 TTTTCCTTATAACAAAGACA 323 TTTAGGGATTTTTTTTTTTCCTTAT 343 _human_6389 AACAAAGACATCACCAGGAT 4 TGFBR1_NM_004612 TGTATTACTTGTTTAATAAT 324 TTTTTATAGTTGTGTTGTATTACTT 344 _human_3611 GTTTAATAATAATCTCTAAT 5 TGFBR1_NM_004612 TTATTGAATCAAAGATTGAG 325 TGCTGAAGATATTTTTTATTGAAT 345 _human_3882 CAAAGATTGAGTTACAATTAT 6 TGFBR1_NM_004612 TTCTTACCTAAGTGGATAAA 326 GTTACAATTATACTTTTCTTACCTA 346 _human_3916 AGTGGATAAAATGTACTTTT 7 TGFBR1_NM_004612 ATGTTGCTCAGTTACTCAAA 327 TAAAGTATGGGTATTATGTTGCTC 347 _human_5559 AGTTACTCAAATGGTACTGTA 8 TGFBR1_NM_004612 ATATTTGTACCCCAAATAAC 328 GGTACTGTATTGTTTATATTTGTA 348 _human_5595 CCCCAAATAACATCGTCTGTA 9 TGFBR1_NM_004612 TGTAAATGTAAACTTCTAAA 329 TTATGCAATCTTGTTTGTAAATGT 349 _human_5222 AAACTTCTAAAAATATGGTTA 10 TGFBR1_NM_004612 AGAATGAGTGACATATTACA 330 AACCAAAGTAATTTTAGAATGAG 350 _human_3435 TGACATATTACATAGGAATTTA 11 TGFBR1_NM_004612 CCATTTCTAAGCCTACCAGA 331 GTTGTTGTTTTTGGGCCATTTCTA 351 _human_3709 AGCCTACCAGATCTGCTTTAT 12 TGFBR1_NM_004612 ATATTCCAAAAGAATGTAAA 332 ATTGTATTTGTAGTAATATTCCAA 352 _human_5826 AAGAATGTAAATAGGAAATAG 13 TGFBR1_NM_004612 TTACTTCCAATGCTATGAAG 333 TATAATAACTGGTTTTTACTTCCA 353 _human_3146 ATGCTATGAAGTCTCTGCAGG 14 TGFBR1_NM_004612 TCTTTATCTGTTCAAAGACT 334 TGTAAGCCATTTTTTTCTTTATCTG 354 _human_2675 TTCAAAGACTTATTTTTTAA 15 TGFBR1_NM_004612 GTCTAAGTATACTTTTAAAA 335 CATTTTAATTGTGTTGTCTAAGTA 355 _human_2529 TACTTTTAAAAAATCAAGTGG 16 TGFBR1_NM_004612 ATCTTTGGACATGTACTGCA 336 GAGATACTAAGGATTATCTTTGG 356 _human_5079 ACATGTACTGCAGCTTCTTGTC 17 TGFBR1_NM_004612 GTGTTGTATTACTTGTTTAA 337 TTTGTTTTTATAGTTGTGTTGTATT 357 _human_3607 ACTTGTTTAATAATAATCTC 18 TGFBR1_NM_004612 TGCTGTAGATGGCAACTAGA 338 CATGCCATATGTAGTTGCTGTAGA 358 _human_5994 TGGCAACTAGAACCTTTGAGT 19 TGFBR1_NM_004612 TCTTTCACTTATTCAGAACA 339 GTATACTATTATTGTTCTTTCACTT 359 _human_2177 ATTCAGAACATTACATGCCT 20 TGFBR1_NM_004612 GTATTTGTAGTAATATTCCA 340 TTTAAATTGTATATTGTATTTGTA 360 _human_5814 GTAATATTCCAAAAGAATGTA Accession: NM_032782 HUGO gene symbol: HAVCR2 1 HAVCR2_NM_032782 CTCATAGCAAAGAGAAGATA 361 TTTTCAAATGGTATTCTCATAGCA 381 _human_937 AAGAGAAGATACAGAATTTAA 2 HAVCR2_NM_032782 GTATTCTCATAGCAAAGAGA 362 TTTAATTTTCAAATGGTATTCTCAT 382 _human_932 AGCAAAGAGAAGATACAGAA 3 HAVCR2_NM_032782 TTGCTTGTTGTGTGCTTGAA 363 TGTATTGGCCAAGTTTTGCTTGTT 383 _human_2126 GTGTGCTTGAAAGAAAATATC 4 HAVCR2_NM_032782 TATTCGTGGACCAAACTGAA 364 TCTGACCAACTTCTGTATTCGTGG 384 _human_2171 ACCAAACTGAAGCTATATTTT 5 HAVCR2_NM_032782 ATTGTGGAGTAGACAGTTGG 365 GCTACTGCTCATGTGATTGTGGA 385 _human_158 GTAGACAGTTGGAAGAAGTACC 6 HAVCR2_NM_032782 GTTGTGTGCTTGAAAGAAAA 366 GGCCAAGTTTTGCTTGTTGTGTGC 386 _human_2132 TTGAAAGAAAATATCTCTGAC 7 HAVCR2_NM_032782 TGTTGTGTGCTTGAAAGAAA 367 TGGCCAAGTTTTGCTTGTTGTGTG 387 _human_2131 CTTGAAAGAAAATATCTCTGA 8 HAVCR2_NM_032782 CCCTAAACTTAAATTTCAAG 368 TTGACAGAGAGTGGTCCCTAAAC 388 _human_2313 TTAAATTTCAAGACGGTATAGG 9 HAVCR2_NM_032782 ACATCCAGATACTGGCTAAA 369 GATGTGAATTATTGGACATCCAG 389 _human_489 ATACTGGCTAAATGGGGATTTC 10 HAVCR2_NM_032782 CATTTTCAGAAGATAATGAC 370 GGAGCAGAGTTTTCCCATTTTCAG 390 _human_1272 AAGATAATGACTCACATGGGA 11 HAVCR2_NM_032782 CACATTGGCCAATGAGTTAC 371 TCTAACACAAATATCCACATTGGC 391 _human_785 CAATGAGTTACGGGACTCTAG 12 HAVCR2_NM_032782 TGCTTGTTGTGTGCTTGAAA 372 GTATTGGCCAAGTTTTGCTTGTTG 392 _human_2127 TGTGCTTGAAAGAAAATATCT 13 HAVCR2_NM_032782 GAGTAGACAGTTGGAAGAAG 373 GCTCATGTGATTGTGGAGTAGAC 393 _human_164 AGTTGGAAGAAGTACCCAGTCC 14 HAVCR2_NM_032782 TTGTTGTGTGCTTGAAAGAA 374 TTGGCCAAGTTTTGCTTGTTGTGT 394 _human_2130 GCTTGAAAGAAAATATCTCTG 15 HAVCR2_NM_032782 CGGCGCTTTAATTTTCAAAT 375 TCTGGCTCTTATCTTCGGCGCTTT 395 _human_911 AATTTTCAAATGGTATTCTCA 16 HAVCR2_NM_032782 TTTGGCACAGAAAGTCTAAA 376 TGAAAGCATAACTTTTTTGGCACA 396 _human_1543 GAAAGTCTAAAGGGGCCACTG 17 HAVCR2_NM_032782 GATCTGTCTTGCTTATTGTT 377 AGACGGTATAGGCTTGATCTGTC 397 _human_2346 TTGCTTATTGTTGCCCCCTGCG 18 HAVCR2_NM_032782 GGTGTGTATTGGCCAAGTTT 378 GAAGTGCATTTGATTGGTGTGTA 398 _human_2107 TTGGCCAAGTTTTGCTTGTTGT 19 HAVCR2_NM_032782 CCCATTTTCAGAAGATAATG 379 ATGGAGCAGAGTTTTCCCATTTTC 399 _human_1270 AGAAGATAATGACTCACATGG 20 HAVCR2_NM_032782 TGGCACAGAAAGTCTAAAGG 380 AAAGCATAACTTTTTTGGCACAGA 400 _human_1545 AAGTCTAAAGGGGCCACTGAT Accession: NM_002987 HUGO gene symbol: CCL17 1 CCL17_NM_002987 AAATACCTGCAAAGCCTTGA 401 GTGAAGAATGCAGTTAAATACCTGC 421 _human_385 AAAGCCTTGAGAGGTCTTGA 2 CCL17_NM_002987 TTTTGTAACTGTGCAGGGCA 402 CAGGGATGCCATCGTTTTTGTAACT 422 _human_318 GTGCAGGGCAGGGCCATCTG 3 CCL17_NM_002987 AGAGTGAAGAATGCAGTTAA 403 GACCCCAACAACAAGAGAGTGAAG 423 _human_367 AATGCAGTTAAATACCTGCAA 4 CCL17_NM_002987 AAGCCTTGAGAGGTCTTGAA 404 AGTTAAATACCTGCAAAGCCTTGAG 424 _human_396 AGGTCTTGAAGCCTCCTCAC 5 CCL17_NM_002987 AATACCTGCAAAGCCTTGAG 405 TGAAGAATGCAGTTAAATACCTGCA 425 _human_386 AAGCCTTGAGAGGTCTTGAA 6 CCL17_NM_002987 TGCAGTTAAATACCTGCAAA 406 CAAGAGAGTGAAGAATGCAGTTAA 426 _human_378 ATACCTGCAAAGCCTTGAGAG 7 CCL17_NM_002987 CAACAACAAGAGAGTGAAGA 407 CATCTGTTCGGACCCCAACAACAAG 427 _human_357 AGAGTGAAGAATGCAGTTAA 8 CCL17_NM_002987 CTGAATTCAAAACCAGGGTG 408 CTGCTGATGGGAGAGCTGAATTCAA 428 _human_55 AACCAGGGTGTCTCCCTGAG 9 CCL17_NM_002987 ATACCTGCAAAGCCTTGAGA 409 GAAGAATGCAGTTAAATACCTGCAA 429 _human_387 AGCCTTGAGAGGTCTTGAAG 10 CCL17_NM_002987 TTCCCCTTAGAAAGCTGAAG 410 ACTTCAAGGGAGCCATTCCCCTTAG 430 _human_254 AAAGCTGAAGACGTGGTACC 11 CCL17_NM_002987 GGAGAGCTGAATTCAAAACC 411 CACCGCCTGCTGATGGGAGAGCTG 431 _human_49 AATTCAAAACCAGGGTGTCTC 12 CCL17_NM_002987 GCAGTTAAATACCTGCAAAG 412 AAGAGAGTGAAGAATGCAGTTAAA 432 _human_379 TACCTGCAAAGCCTTGAGAGG 13 CCL17_NM_002987 GAAGAATGCAGTTAAATACC 413 CAACAACAAGAGAGTGAAGAATGC 433 _human_372 AGTTAAATACCTGCAAAGCCT 14 CCL17_NM_002987 ATGCAGTTAAATACCTGCAA 414 ACAAGAGAGTGAAGAATGCAGTTA 434 _human_377 AATACCTGCAAAGCCTTGAGA 15 CCL17_NM_002987 CATTCCCCTTAGAAAGCTGA 415 GTACTTCAAGGGAGCCATTCCCCTT 435 _human_252 AGAAAGCTGAAGACGTGGTA 16 CCL17_NM_002987 AGAGCTGAATTCAAAACCAG 416 CCGCCTGCTGATGGGAGAGCTGAAT 436 _human_51 TCAAAACCAGGGTGTCTCCC 17 CCL17_NM_002987 GATGGGAGAGCTGAATTCAA 417 GTGTCACCGCCTGCTGATGGGAGA 437 _human_45 GCTGAATTCAAAACCAGGGTG 18 CCL17_NM_002987 TGATGGGAGAGCTGAATTCA 418 AGTGTCACCGCCTGCTGATGGGAGA 438 _human_44 GCTGAATTCAAAACCAGGGT 19 CCL17_NM_002987 ACTTTGAGCTCACAGTGTCA 419 GCTCAGAGAGAAGTGACTTTGAGCT 439 _human_16 CACAGTGTCACCGCCTGCTG 20 CCL17_NM_002987 GAGTGAAGAATGCAGTTAAA 420 ACCCCAACAACAAGAGAGTGAAGA 440 _human_368 ATGCAGTTAAATACCTGCAAA Accession: NM_002990 HUGO gene symbol: CCL22 1 CCL22_NM_002990 GTATTTGAAAACAGAGTAAA 441 GCTGGAGTTATATATGTATTTGAA 461 _human_2083 AACAGAGTAAATACTTAAGAG 2 CCL22_NM_002990 CAATAAGCTGAGCCAATGAA 442 GGTGAAGATGATTCTCAATAAGC 462 _human_298 TGAGCCAATGAAGAGCCTACTC 3 CCL22_NM_002990 TACTTAAGAGGCCAAATAGA 443 TGAAAACAGAGTAAATACTTAAG 463 _human_2103 AGGCCAAATAGATGAATGGAAG 4 CCL22_NM_002990 ATGTATTTGAAAACAGAGTA 444 AAGCTGGAGTTATATATGTATTTG 464 _human_2081 AAAACAGAGTAAATACTTAAG 5 CCL22_NM_002990 TTCATACAGCAAGTATGGGA 445 TTGAGAAATATTCTTTTCATACAG 465 _human_2496 CAAGTATGGGACAGCAGTGTC 6 CCL22_NM_002990 CTGCAGACAAAATCAATAAA 446 GAGCCCAGAAAGTGGCTGCAGAC 466 _human_1052 AAAATCAATAAAACTAATGTCC 7 CCL22_NM_002990 TGCAGACAAAATCAATAAAA 447 AGCCCAGAAAGTGGCTGCAGACA 467 _human_1053 AAATCAATAAAACTAATGTCCC 8 CCL22_NM_002990 GGCCAAATAGATGAATGGAA 448 AGTAAATACTTAAGAGGCCAAAT 468 _human_2112 AGATGAATGGAAGAATTTTAGG 9 CCL22_NM_002990 AATAAGCTGAGCCAATGAAG 449 GTGAAGATGATTCTCAATAAGCT 469 _human_299 GAGCCAATGAAGAGCCTACTCT 10 CCL22_NM_002990 AAGAGGCCAAATAGATGAAT 450 ACAGAGTAAATACTTAAGAGGCC 470 _human_2108 AAATAGATGAATGGAAGAATTT 11 CCL22_NM_002990 AAATAGATGAATGGAAGAAT 451 AATACTTAAGAGGCCAAATAGAT 471 _human_2116 GAATGGAAGAATTTTAGGAACT 12 CCL22_NM_002990 AAACAGAGTAAATACTTAAG 452 TATATATGTATTTGAAAACAGAGT 472 _human_2091 AAATACTTAAGAGGCCAAATA 13 CCL22_NM_002990 AGCTGGAGTTATATATGTAT 453 TGACTTGGTATTATAAGCTGGAG 473 _human_2067 TTATATATGTATTTGAAAACAG 14 CCL22_NM_002990 ACCTTTGACTTGGTATTATA 454 ATGGTGTGAAAGACTACCTTTGA 474 _human_2047 CTTGGTATTATAAGCTGGAGTT 15 CCL22_NM_002990 AACCTTCAGGGATAAGGAGA 455 TGGCGTGGTGTTGCTAACCTTCA 475 _human_238 GGGATAAGGAGATCTGTGCCGA 16 CCL22_NM_002990 GTGAAAGACTACCTTTGACT 456 AATTCATGCTATGGTGTGAAAGA 476 _human_2037 CTACCTTTGACTTGGTATTATA 17 CCL22_NM_002990 CTATGGTGTGAAAGACTACC 457 ACAATCAAATTCATGCTATGGTGT 477 _human_2030 GAAAGACTACCTTTGACTTGG 18 CCL22_NM_002990 CACTACGGCTGGCTAATTTT 458 ATTACAGGTGTGTGCCACTACGG 478 _human_1682 CTGGCTAATTTTTGTATTTTTA 19 CCL22_NM_002990 GGAGTTATATATGTATTTGA 459 TTGGTATTATAAGCTGGAGTTATA 479 _human_2071 TATGTATTTGAAAACAGAGTA 20 CCL22_NM_002990 ATATCAATACAGAGACTCAA 460 CCAAAAGGCAGTTACATATCAAT 480 _human_1111 ACAGAGACTCAAGGTCACTAGA Accession: NM_005618 HUGO gene symbol: DLL1 1 DLL1_NM_005618 CTGTTTTGTTAATGAAGAAA 481 TATTTGAGTTTTTTACTGTTTTGTTA 501 _human_3246 ATGAAGAAATTCCTTTTTA 2 DLL1_NM_005618 TTGTATATAAATGTATTTAT 482 TGTGACTATATTTTTTTGTATATAAA 502 _human_3193 TGTATTTATGGAATATTGT 3 DLL1_NM_005618 TGTTTTGTTAATGAAGAAAT 483 ATTTGAGTTTTTTACTGTTTTGTTAA 503 _human_3247 TGAAGAAATTCCTTTTTAA 4 DLL1_NM_005618 AATTTTGGTAAATATGTACA 484 GTTTTTTATAATTTAAATTTTGGTAA 504 _human_3141 ATATGTACAAAGGCACTTC 5 DLL1_NM_005618 AAATTTTATGAATGACAAAA 485 ATATTTTTCCAAAATAAATTTTATGA 505 _human_3293 ATGACAAAAAAAAAAAAAA 6 DLL1_NM_005618 TTTATGGAATATTGTGCAAA 486 TTGTATATAAATGTATTTATGGAATA 506 _human_3208 TTGTGCAAATGTTATTTGA 7 DLL1_NM_005618 TTACTGTTTTGTTAATGAAG 487 TGTTATTTGAGTTTTTTACTGTTTTGT 507 _human_3243 TAATGAAGAAATTCCTTT 8 DLL1_NM_005618 TTCTTGAATTAGAAACACAA 488 TTATGAGCCAGTCTTTTCTTGAATTA 508 _human_2977 GAAACACAAACACTGCCTT 9 DLL1_NM_005618 CAGTTGCTCTTAAGAGAATA 489 CCGTTGCACTATGGACAGTTGCTCTT 509 _human_2874 AAGAGAATATATATTTAAA 10 DLL1_NM_005618 CAACTTCAAAAGACACCAAG 490 CGGACTCGGGCTGTTCAACTTCAAA 510 _human_2560 AGACACCAAGTACCAGTCGG 11 DLL1_NM_005618 TCCAAAATAAATTTTATGAA 491 TTTTTAAAATATTTTTCCAAAATAAA 511 _human_3285 TTTTATGAATGACAAAAAA 12 DLL1_NM_005618 GAACTGAATTACGCATAAGA 492 TATATTTAAATGGGTGAACTGAATT 512 _human_2909 ACGCATAAGAAGCATGCACT 13 DLL1_NM_005618 GGATTTTGTGACAAACCAGG 493 TGTGATGAGCAGCATGGATTTTGTG 513 _human_1173 ACAAACCAGGGGAATGCAAG 14 DLL1_NM_005618 TACTGTTTTGTTAATGAAGA 494 GTTATTTGAGTTTTTTACTGTTTTGTT 514 _human_3244 AATGAAGAAATTCCTTTT 15 DLL1_NM_005618 TTTGGTAAATATGTACAAAG 495 TTTTATAATTTAAATTTTGGTAAATA 515 _human_3144 TGTACAAAGGCACTTCGGG 16 DLL1_NM_005618 CCAAAATAAATTTTATGAAT 496 TTTTAAAATATTTTTCCAAAATAAAT 516 _human_3286 TTTATGAATGACAAAAAAA 17 DLL1_NM_005618 ATAATTTAAATTTTGGTAAA 497 TGATGTTCGTTTTTTATAATTTAAAT 517 _human_3133 TTTGGTAAATATGTACAAA 18 DLL1_NM_005618 AAATGGGTGAACTGAATTAC 498 AGAGAATATATATTTAAATGGGTGA 518 _human_2901 ACTGAATTACGCATAAGAAG 19 DLL1_NM_005618 TTCGGGTCTATGTGACTATA 499 TATGTACAAAGGCACTTCGGGTCTA 519 _human_3168 TGTGACTATATTTTTTTGTA 20 DLL1_NM_005618 ACTGTTTTGTTAATGAAGAA 500 TTATTTGAGTTTTTTACTGTTTTGTTA 520 _human_3245 ATGAAGAAATTCCTTTTT Accession: NM_000639 HUGO gene symbol: FASLG 1 FASLG_NM_000639 TAGCTCCTCAACTCACCTAA 521 GGTTCAAAATGTCTGTAGCTCCTC 541 _human_1154 AACTCACCTAATGTTTATGAG 2 FASLG_NM_000639 ATGTTTTCCTATAATATAAT 522 TGTCAGCTACTAATGATGTTTTCC 542 _human_1771 TATAATATAATAAATATTTAT 3 FASLG_NM_000639 TTTTCCTATAATATAATAAA 523 CAGCTACTAATGATGTTTTCCTAT 543 _human_1774 AATATAATAAATATTTATGTA 4 FASLG_NM_000639 TTCCTATAATATAATAAATA 524 GCTACTAATGATGTTTTCCTATAA 544 _human_1776 TATAATAAATATTTATGTAGA 5 FASLG_NM_000639 TGCATTTGAGGTCAAGTAAG 525 GAGGGTCTTCTTACATGCATTTGA 545 _human_1086 GGTCAAGTAAGAAGACATGAA 6 FASLG_NM_000639 ATTGATTGTCAGCTACTAAT 526 TAGTGCTTAAAAATCATTGATTGT 546 _human_1750 CAGCTACTAATGATGTTTTCC 7 FASLG_NM_000639 AAATGAAAACATGTAATAAA 527 ATGTGCATTTTTGTGAAATGAAAA 547 _human_1820 CATGTAATAAAAAGTATATGT 8 FASLG_NM_000639 ATTGTGAAGTACATATTAGG 528 AGAGAGAATGTAGATATTGTGAA 548 _human_1659 GTACATATTAGGAAAATATGGG 9 FASLG_NM_000639 GCTTTCTGGAGTGAAGTATA 529 CTATGGAATTGTCCTGCTTTCTGG 549 _human_667 AGTGAAGTATAAGAAGGGTGG 10 FASLG_NM_000639 CATTTGGTCAAGATTTTGAA 530 GGAAAATATGGGTTGCATTTGGT 550 _human_1692 CAAGATTTTGAATGCTTCCTGA 11 FASLG_NM_000639 GGCTTATATAAGCTCTAAGA 531 TCTCAGACGTTTTTCGGCTTATAT 551 _human_986 AAGCTCTAAGAGAAGCACTTT 12 FASLG_NM_000639 ACCAGTGCTGATCATTTATA 532 GCAGTGTTCAATCTTACCAGTGCT 552 _human_911 GATCATTTATATGTCAACGTA 13 FASLG_NM_000639 CCATTTAACAGGCAAGTCCA 533 GCTGAGGAAAGTGGCCCATTTAA 553 _human_598 CAGGCAAGTCCAACTCAAGGTC 14 FASLG_NM_000639 AAGTACATATTAGGAAAATA 534 AATGTAGATATTGTGAAGTACAT 554 _human_1665 ATTAGGAAAATATGGGTTGCAT 15 FASLG_NM_000639 TGTGTGTGTGTATGACTAAA 535 GTGTGTGTGTGTGTGTGTGTGTG 555 _human_1625 TGTATGACTAAAGAGAGAATGT 16 FASLG_NM_000639 AAGAGGGAGAAGCATGAAAA 536 CTGGGCTGCCATGTGAAGAGGGA 556 _human_1238 GAAGCATGAAAAAGCAGCTACC 17 FASLG_NM_000639 GTGTATGACTAAAGAGAGAA 537 TGTGTGTGTGTGTGTGTGTATGA 557 _human_1632 CTAAAGAGAGAATGTAGATATT 18 FASLG_NM_000639 GTATTTCCAGTGCAATTGTA 538 CCTAACACAGCATGTGTATTTCCA 558 _human_1581 GTGCAATTGTAGGGGTGTGTG 19 FASLG_NM_000639 CAACTCTAATAGTGCTTAAA 539 ATGCTTCCTGACAATCAACTCTAA 559 _human_1726 TAGTGCTTAAAAATCATTGAT 20 FASLG_NM_000639 GTGTGTGTGTATGACTAAAG 540 TGTGTGTGTGTGTGTGTGTGTGT 560 human_1626 GTATGACTAAAGAGAGAATGTA Accession: NM_001267706 HUGO gene symbol: CD274 1 CD274_NM_001267706 ACCTGCATTAATTTAATAAA 561 ATTGTCACTTTTTGTACCTGCATTA 581 _human_3222 ATTTAATAAAATATTCTTAT 2 CD274_NM_001267706 AACTTGCCCAAACCAGTAAA 562 GCAAACAGATTAAGTAACTTGCC 582 _human_1538 CAAACCAGTAAATAGCAGACCT 3 CD274_NM_001267706 ATTTGCTCACATCTAGTAAA 563 ACTTGCTGCTTAATGATTTGCTCA 583 _human_1218 CATCTAGTAAAACATGGAGTA 4 CD274_NM_001267706 CCTTTGCCATATAATCTAAT 564 TTTATTCCTGATTTGCCTTTGCCAT 584 _human_1998 ATAATCTAATGCTTGTTTAT 5 CD274_NM_001267706 ATATAGCAGATGGAATGAAT 565 ATTTTAGTGTTTCTTATATAGCAG 585 _human_2346 ATGGAATGAATTTGAAGTTCC 6 CD274_NM_001267706 GCCTTTGCCATATAATCTAA 566 ATTTATTCCTGATTTGCCTTTGCCA 586 _human_1997 TATAATCTAATGCTTGTTTA 7 CD274_NM_001267706 GATTTGCCTTTGCCATATAA 567 ATTATATTTATTCCTGATTTGCCTT 587 _human_1992 TGCCATATAATCTAATGCTT 8 CD274_NM_001267706 AATTTTCATTTACAAAGAGA 568 CTTAATAATCAGAGTAATTTTCAT 588 _human_1905 TTACAAAGAGAGGTCGGTACT 9 CD274_NM_001267706 AGTGTTTCTTATATAGCAGA 569 ATTTTTATTTATTTTAGTGTTTCTT 589 _human_2336 ATATAGCAGATGGAATGAAT 10 CD274_NM_001267706 GCTTTCTGTCAAGTATAAAC 570 GAACTTTTGTTTTCTGCTTTCTGTC 590 _human_2656 AAGTATAAACTTCACTTTGA 11 CD274_NM_001267706 CATTTGGAAATGTATGTTAA 571 TCTAAAGATAGTCTACATTTGGAA 591 _human_2235 ATGTATGTTAAAAGCACGTAT 12 CD274_NM_001267706 TTATTTTAGTGTTTCTTATA 572 CTTTGCTATTTTTATTTATTTTAGT 592 _human_2329 GTTTCTTATATAGCAGATGG 13 CD274_NM_001267706 GTGGTAGCCTACACACATAA 573 CAGCTTTACAATTATGTGGTAGCC 593 _human_1433 TACACACATAATCTCATTTCA 14 CD274_NM_001267706 ATGAGGAGATTAACAAGAAA 574 GGAGCTCATAGTATAATGAGGAG 594 _human_1745 ATTAACAAGAAAATGTATTATT 15 CD274_NM_001267706 CAATTTTGTCGCCAAACTAA 575 TTGTAGTAGATGTTACAATTTTGT 595 _human_1183 CGCCAAACTAAACTTGCTGCT 16 CD274_NM_001267706 TATATAGCAGATGGAATGAA 576 TATTTTAGTGTTTCTTATATAGCA 596 _human_2345 GATGGAATGAATTTGAAGTTC 17 CD274_NM_001267706 AAATGCCACTAAATTTTAAA 577 CTGTCTTTTCTATTTAAATGCCACT 597 _human_2069 AAATTTTAAATTCATACCTT 18 CD274_NM_001267706 TCTTTCCCATAGCTTTTCAT 578 TTTGTTTCTAAGTTATCTTTCCCAT 598 _human_2414 AGCTTTTCATTATCTTTCAT 19 CD274_NM_001267706 TATATTCATGACCTACTGGC 579 GATATTTGCTGTCTTTATATTCAT 599 _human_129 GACCTACTGGCATTTGCTGAA 20 CD274_NM_001267706 GTCCAGTGTCATAGCATAAG 580 TATTATTACAATTTAGTCCAGTGT 600 _human_1783 CATAGCATAAGGATGATGCGA Accession: NM_002164 HUGO gene symbol: IDO1 1 IDO1_NM_002164 ATTCTGTCATAATAAATAAA 601 AAAAAAAAAAGATATATTCTGTCA 621 _human_1896 TAATAAATAAAAATGCATAAG 2 IDO1_NM_002164 TATCTTATCATTGGAATAAA 602 AAGTTTTGTAATCTGTATCTTATCA 622 _human_1532 TTGGAATAAAATGACATTCA 3 IDO1_NM_002164 GTGATGGAGACTGCAGTAAA 603 TTTTGTTCTCATTTCGTGATGGAGA 623 _human_578 CTGCAGTAAAGGATTCTTCC 4 IDO1_NM_002164 TTCTGTCATAATAAATAAAA 604 AAAAAAAAAGATATATTCTGTCAT 624 _human_1897 AATAAATAAAAATGCATAAGA 5 IDO1_NM_002164 CTTGTAGGAAAACAACAAAA 605 AATACCTGTGCATTTCTTGTAGGAA 625 _human_1473 AACAACAAAAGGTAATTATG 6 IDO1_NM_002164 ATAAAATGACATTCAATAAA 606 TATCTTATCATTGGAATAAAATGAC 626 _human_1547 ATTCAATAAATAAAAATGCA 7 IDO1_NM_002164 CGTAAGGTCTTGCCAAGAAA 607 GGTCATGGAGATGTCCGTAAGGTC 627 _human_412 TTGCCAAGAAATATTGCTGTT 8 IDO1_NM_002164 TCTTGTAGGAAAACAACAAA 608 AAATACCTGTGCATTTCTTGTAGGA 628 _human_1472 AAACAACAAAAGGTAATTAT 9 IDO1_NM_002164 AACTGGAGGCACTGATTTAA 609 ACTGGAAGCCAAAGGAACTGGAG 629 _human_1248 GCACTGATTTAATGAATTTCCT 10 IDO1_NM_002164 CAATACAAAAGACCTCAAAA 610 GTTTTACCAATAATGCAATACAAAA 630 _human_1440 GACCTCAAAATACCTGTGCA 11 IDO1_NM_002164 TGCTTCTGCAATCAAAGTAA 611 GGTGGAAATAGCAGCTGCTTCTGC 631 _human_636 AATCAAAGTAATTCCTACTGT 12 IDO1_NM_002164 AATGACATTCAATAAATAAA 612 TTATCATTGGAATAAAATGACATTC 632 _human_1551 AATAAATAAAAATGCATAAG 13 IDO1_NM_002164 ATCATTGGAATAAAATGACA 613 TGTAATCTGTATCTTATCATTGGAA 633 _human_1538 TAAAATGACATTCAATAAAT 14 IDO1_NM_002164 ACCAATAATGCAATACAAAA 614 ACTATGCAATGTTTTACCAATAATG 634 _human_1430 CAATACAAAAGACCTCAAAA 15 IDO1_NM_002164 ATCTGTATCTTATCATTGGA 615 ACTAGAAGTTTTGTAATCTGTATCT 635 _human_1527 TATCATTGGAATAAAATGAC 16 IDO1_NM_002164 ATCTTATCATTGGAATAAAA 616 AGTTTTGTAATCTGTATCTTATCAT 636 _human_1533 TGGAATAAAATGACATTCAA 17 IDO1_NM_002164 CAGCTGCTTCTGCAATCAAA 617 TATTGGTGGAAATAGCAGCTGCTT 637 _human_632 CTGCAATCAAAGTAATTCCTA 18 IDO1_NM_002164 GCAATACAAAAGACCTCAAA 618 TGTTTTACCAATAATGCAATACAAA 638 _human_1439 AGACCTCAAAATACCTGTGC 19 IDO1_NM_002164 TCCTACTGTATTCAAGGCAA 619 TGCAATCAAAGTAATTCCTACTGTA 639 _human_657 TTCAAGGCAATGCAAATGCA 20 IDO1_NM_002164 CAGAGCCACAAACTAATACT 620 CATTACCCATTGTAACAGAGCCAC 640 _human_1398 AAACTAATACTATGCAATGTT Accession: NM_001558 HUGO gene symbol: IL10RA 1 IL10RA_NM_001558_ TTGTTCATTTATTTATTGGA 641 CTTTATTTATTTATTTTGTTCATTT 661 human_3364 ATTTATTGGAGAGGCAGCAT 2 IL10RA_NM_001558_ TTATTCCAATAAATTGTCAA 642 AGTGATACATGTTTTTTATTCCAA 662 human_3626 TAAATTGTCAAGACCACAGGA 3 IL10RA_NM_001558_ TATTTTCTGGACACTCAAAC 643 AGATCTTAAGGTATATATTTTCTG 663 human_2395 GACACTCAAACACATCATAAT 4 IL10RA_NM_001558_ TTTATTGGAGAGGCAGCATT 644 TATTTTGTTCATTTATTTATTGGAG 664 human_3375 AGGCAGCATTGCACAGTGAA 5 IL10RA_NM_001558_ ACCTTGGAGAAGTCACTTAT 645 GTTTCCAGTGGTATGACCTTGGA 665 human_3469 GAAGTCACTTATCCTCTTGGAG 6 IL10RA_NM_001558_ TTATTTATTTATTTTGTTCA 646 GTTCCCTTGAAAGCTTTATTTATTT 666 human_3351 ATTTTGTTCATTTATTTATT 7 IL10RA_NM_001558_ CTCTTTCCTGTATCATAAAG 647 TCTCCCTCCTAGGAACTCTTTCCT 667 human_2108 GTATCATAAAGGATTATTTGC 8 IL10RA_NM_001558_ CTGAGGAAATGGGTATGAAT 648 GGATGTGAGGTTCTGCTGAGGAA 668 human_3563 ATGGGTATGAATGTGCCTTGAA 9 IL10RA_NM_001558_ GAATGTGCCTTGAACACAAA 649 TGAGGAAATGGGTATGAATGTGC 669 human_3579 CTTGAACACAAAGCTCTGTCAA 10 IL10RA_NM_001558_ GGACACTCAAACACATCATA 650 AGGTATATATTTTCTGGACACTCA 670 human_2403 AACACATCATAATGGATTCAC 11 IL10RA_NM_001558_ CTGTATCATAAAGGATTATT 651 CCTAGGAACTCTTTCCTGTATCAT 671 human_2115 AAAGGATTATTTGCTCAGGGG 12 IL10RA_NM_001558_ TCACTTCCGAGAGTATGAGA 652 TGAAAGCATCTTCAGTCACTTCCG 672 human_563 AGAGTATGAGATTGCCATTCG 13 IL10RA_NM_001558_ TCTCTGGAGCATTCTGAAAA 653 TCTCAGCCCTGCCTTTCTCTGGAG673 human_3197 CATTCTGAAAACAGATATTCT 14 IL10RA_NM_001558_ TTATGCCAGAGGCTAACAGA 654 AAGCTGGCTTGTTTCTTATGCCAG 674 human_2987 AGGCTAACAGATCCAATGGGA 15 IL10RA_NM_001558_ AGTGGCATTGACTTAGTTCA 655 AGGGGCCAGGATGACAGTGGCA 675 human_1278 TTGACTTAGTTCAAAACTCTGAG 16 IL10RA_NM_001558_ TTTCTGGACACTCAAACACA 656 TCTTAAGGTATATATTTTCTGGAC 676 human_2398 ACTCAAACACATCATAATGGA 17 IL10RA_NM_001558_ GCATTGCACAGTGAAAGAAT 657 TTTATTGGAGAGGCAGCATTGCA 677 human_3390 CAGTGAAAGAATTCTGGATATC 18 IL10RA_NM_001558_ GACCTTGGAGAAGTCACTTA 658 TGTTTCCAGTGGTATGACCTTGGA 678 human_3468 GAAGTCACTTATCCTCTTGGA 19 IL10RA_NM_001558_ TCACGTTCACACACAAGAAA 659 AGGTGCCGGGAAACTTCACGTTC 679 human_610 ACACACAAGAAAGTAAAACATG 20 IL10RA_NM_001558_ ACTTTGCTGTTTCCAGTGGT 660 GAAATTCTAGCTCTGACTTTGCTG 680 human_3446 TTTCCAGTGGTATGACCTTGG Accession: NM_000214 HUGO gene symbol: JAG1 1 JAG1_NM_000214 TATTTGATTTATTAACTTAA 681 ATTAATCACTGTGTATATTTGATTT 701 _human_4799 ATTAACTTAATAATCAAGAG 2 JAG1_NM_000214 GAAAAGTAATATTTATTAAA 682 TTGGCAATAAATTTTGAAAAGTAA 702 _human_5658 TATTTATTAAATTTTTTTGTA 3 JAG1_NM_000214 ACTTTGTATAGTTATGTAAA 683 AATGTCAAAAGTAGAACTTTGTAT 703 _human_4752 AGTTATGTAAATAATTCTTTT 4 JAG1_NM_000214 GAATACTTGAACCATAAAAT 684 TCTAATAAGCTAGTTGAATACTTGA 704 _human_5418 ACCATAAAATGTCCAGTAAG 5 JAG1_NM_000214 TCTTGGCAATAAATTTTGAA 685 TCTTTGATGTGTTGTTCTTGGCAAT 705 _human_5641 AAATTTTGAAAAGTAATATT 6 JAG1_NM_000214 TTTCTGCTTTAGACTTGAAA 686 TGTTTGTTTTTTGTTTTTCTGCTTTA 706 _human_5150 GACTTGAAAAGAGACAGGC 7 JAG1_NM_000214 TATATTTATTGACTCTTGAG 687 GATCATAGTTTTATTTATATTTATT 707 _human_4526 GACTCTTGAGTTGTTTTTGT 8 JAG1_NM_000214 TATGATGACGTACAAGTAGT 688 TTTGTATATTGGTTTTATGATGACG 708 _human_4566 TACAAGTAGTTCTGTATTTG 9 JAG1_NM_000214 GTGTTGTTCTTGGCAATAAA 689 AAATGCATCTTTGATGTGTTGTTCT 709 _human_5634 TGGCAATAAATTTTGAAAAG 10 JAG1_NM_000214 CTGATCTAAAAGGGAATAAA 690 CCTTTTTCCATGCAGCTGATCTAAA 710 _human_173 AGGGAATAAAAGGCTGCGCA 11 JAG1_NM_000214 TACGACGTCAGATGTTTAAA 691 GATGGAATTTTTTTGTACGACGTCA 711 _human_5031 GATGTTTAAAACACCTTCTA 12 JAG1_NM_000214 AATAATCAAGAGCCTTAAAA 692 TTGATTTATTAACTTAATAATCAAG 712 _human_4817 AGCCTTAAAACATCATTCCT 13 JAG1_NM_000214 GTATGAAAACATGGAACAGT 693 TTATTAAATTTTTTTGTATGAAAAC 713 _human_5685 ATGGAACAGTGTGGCCTCTT 14 JAG1_NM_000214 TGGTTTTATGATGACGTACA 694 GTTGTTTTTGTATATTGGTTTTATG 714 _human_4560 ATGACGTACAAGTAGTTCTG 15 JAG1_NM_000214 TTCTGCTTTAGACTTGAAAA 695 GTTTGTTTTTTGTTTTTCTGCTTTAG 715 _human_5151 ACTTGAAAAGAGACAGGCA 16 JAG1_NM_000214 CTTGGCAATAAATTTTGAAA 696 CTTTGATGTGTTGTTCTTGGCAATA 716 _human_5642 AATTTTGAAAAGTAATATTT 17 JAG1_NM_000214 TTTAATCTACTGCATTTAGG 697 GATTTGATTTTTTTTTTTAATCTACT 717 _human_5377 GCATTTAGGGAGTATTCTA 18 JAG1_NM_000214 TGTATAGTTATGTAAATAAT 698 TCAAAAGTAGAACTTTGTATAGTTA 718 _human_4756 TGTAAATAATTCTTTTTTAT 19 JAG1_NM_000214 ATTTATATTTATTGACTCTT 699 TTAGATCATAGTTTTATTTATATTTA 719 _human_4523 TTGACTCTTGAGTTGTTTT 20 JAG1_NM_000214 CTTTTCACCATTCGTACATA 700 TGTAAATTCTGATTTCTTTTCACCAT 720 _human_5325 TCGTACATAATACTGAACC Accession: NM_002226 HUGO gene symbol: JAG2 1 JAG2_NM_002226 CGTTTCTTTAACCTTGTATA 721 AATGTTTATTTTCTACGTTTCTTTAA 741 _human_4266 CCTTGTATAAATTATTCAG 2 JAG2_NM_002226 TAAATGAATGAACGAATAAA 722 GGCAGAACAAATGAATAAATGAAT 742 _human_5800 GAACGAATAAAAATTTTGACC 3 JAG2_NM_002226 TCATTCATTTATTCCTTTGT 723 GGTCAAAATTTTTATTCATTCATTT 743 _human_5450 ATTCCTTTGTTTTGCTTGGT 4 JAG2_NM_002226 GTAAATGTGTACATATTAAA 724 TGAAAGTGCATTTTTGTAAATGTGT 744 _human_5021 ACATATTAAAGGAAGCACTC 5 JAG2_NM_002226 ACCCACGAATACGTATCAAG 725 AGTATAAAATTGCTTACCCACGAAT 745 _human_5398 ACGTATCAAGGTCTTAAGGA 6 JAG2_NM_002226 GTTTTATAAAATAGTATAAA 726 AAACAGCTGAAAACAGTTTTATAA 746 _human_5371 AATAGTATAAAATTGCTTACC 7 JAG2_NM_002226 CAACTGAGTCAAGGAGCAAA 727 TGAGGGGTAGGAGGTCAACTGAG 747 _human_5691 TCAAGGAGCAAAGCCAAGAACC 8 JAG2_NM_002226 ATGTGTACATATTAAAGGAA 728 AGTGCATTTTTGTAAATGTGTACAT 748 _human_5025 ATTAAAGGAAGCACTCTGTA 9 JAG2_NM_002226 TTCTTTAACCTTGTATAAAT 729 GTTTATTTTCTACGTTTCTTTAACCT 749 _human_4269 TGTATAAATTATTCAGTAA 10 JAG2_NM_002226 ATTTTCTACGTTTCTTTAAC 730 AAAAACCAAATGTTTATTTTCTACG 750 _human_4258 TTTCTTTAACCTTGTATAAA 11 JAG2_NM_002226 CAGTTTTATAAAATAGTATA 731 TAAAACAGCTGAAAACAGTTTTAT 751 _human_5369 AAAATAGTATAAAATTGCTTA 12 JAG2_NM_002226 GCACAGGCAGAACAAATGAA 732 GAGTGAGGCTGCCTTGCACAGGCA 752 _human_5780 GAACAAATGAATAAATGAATG 13 JAG2_NM_002226 TCAGGCTGAAAACAATGGAG 733 ATTATTCAGTAACTGTCAGGCTGA 753 _human_4302 AAACAATGGAGTATTCTCGGA 14 JAG2_NM_002226 TAAAATTGCTTACCCACGAA 734 TTTTATAAAATAGTATAAAATTGCT 754 _human_5387 TACCCACGAATACGTATCAA 15 JAG2_NM_002226 GTCAGGCTGAAAACAATGGA 735 AATTATTCAGTAACTGTCAGGCTG 755 _human_4301 AAAACAATGGAGTATTCTCGG 16 JAG2_NM_002226 AAATGTGTACATATTAAAGG 736 AAAGTGCATTTTTGTAAATGTGTAC 756 _human_5023 ATATTAAAGGAAGCACTCTG 17 JAG2_NM_002226 CAGTAACTGTCAGGCTGAAA 737 CTTGTATAAATTATTCAGTAACTGT 757 _human_4293 CAGGCTGAAAACAATGGAGT 18 JAG2_NM_002226 GTATTCTCGGATAGTTGCTA 738 GCTGAAAACAATGGAGTATTCTCG 758 _human_4321 GATAGTTGCTATTTTTGTAAA 19 JAG2_NM_002226 TCTCACACAAATTCACCAAA 739 AGGCGGAGAAGTTCCTCTCACACA 759 _human_3994 AATTCACCAAAGATCCTGGCC 20 JAG2_NM_002226 TTGTTTTGCTTGGTCATTCA 740 CATTCATTTATTCCTTTGTTTTGCTT 760 _human_5466 GGTCATTCAGAGGCAAGGT Accession: NM_001315 HUGO gene symbol: MAPK14 1 MAPK14_NM_001315 TCATGCGAAAAGAACCTACA 761 ATTTCAGTCCATCATTCATGCGAAAA 781 _human_670 GAACCTACAGAGAACTGCG 2 MAPK14_NM_001315 AAATGTCAGAAGCTTACAGA 762 CTGAACAACATTGTGAAATGTCAGA 782 _human_833 AGCTTACAGATGACCATGTT 3 MAPK14_NM_001315 AAACATATGAAACATGAAAA 763 GAACTGCGGTTACTTAAACATATGA 783 _human_707 AACATGAAAATGTGATTGGT 4 MAPK14_NM_001315 CAGTTCCTTATCTACCAAAT 764 ACAGATGACCATGTTCAGTTCCTTAT 784 _human_863 CTACCAAATTCTCCGAGGT 5 MAPK14_NM_001315 TCCTGGTACAGACCATATTA 765 TGGAAGAACATTGTTTCCTGGTACA 785 _human_1150 GACCATATTAACCAGCTTCA 6 MAPK14_NM_001315 TTCCTTATCTACCAAATTCT 766 GATGACCATGTTCAGTTCCTTATCTA 786 _human_866 CCAAATTCTCCGAGGTCTA 7 MAPK14_NM_001315 TTCCTGGTACAGACCATATT 767 CTGGAAGAACATTGTTTCCTGGTAC 787 _human_1149 AGACCATATTAACCAGCTTC 8 MAPK14_NM_001315 AAGTATATACATTCAGCTGA 768 ATTCTCCGAGGTCTAAAGTATATAC 788 _human_896 ATTCAGCTGACATAATTCAC 9 MAPK14_NM_001315 CATTACAACCAGACAGTTGA 769 ATGCTGAACTGGATGCATTACAACC 789 _human_1076 AGACAGTTGATATTTGGTCA 10 MAPK14_NM_001315 AGGGACCTAAAACCTAGTAA 770 GCTGACATAATTCACAGGGACCTAA 790 _human_926 AACCTAGTAATCTAGCTGTG 11 MAPK14_NM_001315 CTCTGGAGGAATTCAATGAT 771 TTACACCTGCAAGGTCTCTGGAGGA 791 _human_765 ATTCAATGATGTGTATCTGG 12 MAPK14_NM_001315 TAAACATATGAAACATGAAA 772 AGAACTGCGGTTACTTAAACATATG 792 _human_706 AAACATGAAAATGTGATTGG 13 MAPK14_NM_001315 GATCTGAACAACATTGTGAA 773 CATCTCATGGGGGCAGATCTGAACA 793 _human_815 ACATTGTGAAATGTCAGAAG 14 MAPK14_NM_001315 TCAGTTCCTTATCTACCAAA 774 TACAGATGACCATGTTCAGTTCCTTA 794 _human_862 TCTACCAAATTCTCCGAGG 15 MAPK14_NM_001315 ATAATTCACAGGGACCTAAA 775 ATACATTCAGCTGACATAATTCACA 795 _human_917 GGGACCTAAAACCTAGTAAT 16 MAPK14_NM_001315 CGAGGTCTAAAGTATATACA 776 ATCTACCAAATTCTCCGAGGTCTAA 796 _human_887 AGTATATACATTCAGCTGAC 17 MAPK14_NM_001315 GAAATGTCAGAAGCTTACAG 777 TCTGAACAACATTGTGAAATGTCAG 797 _human_832 AAGCTTACAGATGACCATGT 18 MAPK14_NM_001315 AGCTGTTGACTGGAAGAACA 778 GATGCATAATGGCCGAGCTGTTGAC 798 _human_1125 TGGAAGAACATTGTTTCCTG 19 MAPK14_NM_001315 AAATTCTCCGAGGTCTAAAG 779 AGTTCCTTATCTACCAAATTCTCCGA 799 _human_879 GGTCTAAAGTATATACATT 20 MAPK14_NM_001315 AATGTGATTGGTCTGTTGGA 780 CATATGAAACATGAAAATGTGATTG 800 _human_725 GTCTGTTGGACGTTTTTACA Accession: NM_003745 HUGO gene symbol: SOCS1 1 SOCS1_NM_003745 CTGCTGTGCAGAATCCTATT 801 TCTGGCTTTATTTTTCTGCTGTGCAGAA 821 _human_1141 TCCTATTTTATATTTTT 2 SOCS1_NM_003745 GCTGTGCAGAATCCTATTTT 802 TGGCTTTATTTTTCTGCTGTGCAGAATC 822 _human_1143 CTATTTTATATTTTTTA 3 SOCS1_NM_003745 TTAAAGTCAGTTTAGGTAAT 803 CCTATTTTATATTTTTTAAAGTCAGTTT 823 _human_1170 AGGTAATAAACTTTATT 4 SOCS1_NM_003745 CTGTGCAGAATCCTATTTTA 804 GGCTTTATTTTTCTGCTGTGCAGAATCC 824 _human_1144 TATTTTATATTTTTTAA 5 SOCS1_NM_003745 GTTTACATATACCCAGTATC 805 CTCCTACCTCTTCATGTTTACATATACC 825 _human_1076 CAGTATCTTTGCACAAA 6 SOCS1_NM_003745 ATTTTGTTATTACTTGCCTG 806 CTGGGATGCCGTGTTATTTTGTTATTA 826 _human_837 CTTGCCTGGAACCATGTG 7 SOCS1_NM_003745 TAACTGGGATGCCGTGTTAT 807 CCGTGCACGCAGCATTAACTGGGATG 827 _human_819 CCGTGTTATTTTGTTATTA 8 SOCS1_NM_003745 TGTTATTACTTGCCTGGAAC 808 GATGCCGTGTTATTTTGTTATTACTTGC 828 _human_841 CTGGAACCATGTGGGTA 9 SOCS1_NM_003745 TTTCTGCTGTGCAGAATCCT 809 GTCTCTGGCTTTATTTTTCTGCTGTGCA 829 _human_1138 GAATCCTATTTTATATT 10 SOCS1_NM_003745 CGTGTTATTTTGTTATTACT 810 CATTAACTGGGATGCCGTGTTATTTTG 830 _human_831 TTATTACTTGCCTGGAAC 11 SOCS1_NM_003745 TTTTAAAGTCAGTTTAGGTA 811 ATCCTATTTTATATTTTTTAAAGTCAGT 831 _human_1168 TTAGGTAATAAACTTTA 12 SOCS1_NM_003745 TGCTGTGCAGAATCCTATTT 812 CTGGCTTTATTTTTCTGCTGTGCAGAAT 832 _human_1142 CCTATTTTATATTTTTT 13 SOCS1_NM_003745 GGATGCCGTGTTATTTTGTT 813 ACGCAGCATTAACTGGGATGCCGTGTT 833 _human_825 ATTTTGTTATTACTTGCC 14 SOCS1_NM_003745 TTTAAAGTCAGTTTAGGTAA 814 TCCTATTTTATATTTTTTAAAGTCAGTT 834 _human_1169 TAGGTAATAAACTTTAT 15 SOCS1_NM_003745 TAAAGTCAGTTTAGGTAATA 815 CTATTTTATATTTTTTAAAGTCAGTTTA 835 _human_1171 GGTAATAAACTTTATTA 16 SOCS1_NM_003745 TCTGCTGTGCAGAATCCTAT 816 CTCTGGCTTTATTTTTCTGCTGTGCAGA 836 _human_1140 ATCCTATTTTATATTTT 17 SOCS1_NM_003745 ATATACCCAGTATCTTTGCA 817 CCTCTTCATGTTTACATATACCCAGTAT 837 _human_1082 CTTTGCACAAACCAGGG 18 SOCS1_NM_003745 AGAATCCTATTTTATATTTT 818 ATTTTTCTGCTGTGCAGAATCCTATTTT 838 _human_1150 ATATTTTTTAAAGTCAG 19 SOCS1_NM_003745 GGTTGTTGTAGCAGCTTAAC 819 CCTCTGGGTCCCCCTGGTTGTTGTAGC 839 _human_1011 AGCTTAACTGTATCTGGA 20 SOCS1_NM_003745 CCCAGTATCTTTGCACAAAC 820 TCATGTTTACATATACCCAGTAT 840 _human_1087 CTTTGCACAAACCAGGGGTTGG Accession: NM_003150 HUGO gene symbol: STAT3 1 STAT3_NM_003150 ATATTGCTGTATCTACTTTA 841 TTTTTTTTTTTTGGTATATTGCTGT 861 _human_4897 ATCTACTTTAACTTCCAGAA 2 STAT3_NM_003150 TGTTTGTTAAATCAAATTAG 842 GTTTCTGTGGAATTCTGTTTGTTA 862 _human_4325 AATCAAATTAGCTGGTCTCTG 3 STAT3_NM_003150 TTTATCTAAATGCAAATAAG 843 TGTGGGTGATCTGCTTTTATCTAA 863 _human_2730 ATGCAAATAAGGATGTGTTCT 4 STAT3_NM_003150 ATTTTCCTTTGTAATGTATT 844 TTTATAAATAGACTTATTTTCCTTT 864 _human_3615 GTAATGTATTGGCCTTTTAG 5 STAT3_NM_003150 TATCAGCACAATCTACGAAG 845 GAGTCGAATGTTCTCTATCAGCAC 865 _human_453 AATCTACGAAGAATCAAGCAG 6 STAT3_NM_003150 AGCTTAACTGATAAACAGAA 846 CTTCAGTACATAATAAGCTTAACT 866 _human_4477 GATAAACAGAATATTTAGAAA 7 STAT3_NM_003150 GTTGTTGTTGTTCTTAGACA 847 CAGCTTTTTGTTATTGTTGTTGTTG 867 _human_2870 TTCTTAGACAAGTGCCTCCT 8 STAT3_NM_003150 GTTGTTGTTCTTAGACAAGT 848 CTTTTTGTTATTGTTGTTGTTGTTC 868 _human_2873 TTAGACAAGTGCCTCCTGGT 9 STAT3_NM_003150 TCTGTATTTAAGAAACTTAA 849 TATCAGCATAGCCTTTCTGTATTT 869 _human_3096 AAGAAACTTAAGCAGCCGGGC 10 STAT3_NM_003150 TTATTTTCCTTTGTAATGTA 850 TTTTTATAAATAGACTTATTTTCCT 870 _human_3613 TTGTAATGTATTGGCCTTTT 11 STAT3_NM_003150 TAACTGATAAACAGAATATT 851 AGTACATAATAAGCTTAACTGATA 871 _human_4481 AACAGAATATTTAGAAAGGTG 12 STAT3_NM_003150 ACATTCTGGGCACAAACACA 852 GATCCCGGAAATTTAACATTCTGG 872 _human_1372 GCACAAACACAAAAGTGATGA 13 STAT3_NM_003150 GTGATCTGCTTTTATCTAAA 853 AATGAGTGAATGTGGGTGATCTG 873 _human_2720 CTTTTATCTAAATGCAAATAAG 14 STAT3_NM_003150 CAGACCCGTCAACAAATTAA 854 GCAGAATCTCAACTTCAGACCCGT 874 _human_1044 CAACAAATTAAGAAACTGGAG 15 STAT3_NM_003150 GGAGCTGTTTAGAAACTTAA 855 GGAGGAGAGAATCGTGGAGCTG 875 _human_1148 TTTAGAAACTTAATGAAAAGTGC 16 STAT3_NM_003150 ACCATTGGGTTTAAATCATA 856 GTGAGACTTGGGCTTACCATTGG 876 _human_4523 GTTTAAATCATAGGGACCTAGG 17 STAT3_NM_003150 GGAGAATCTAAGCATTTTAG 857 AATAGGAAGGTTTAAGGAGAATC 877 _human_3573 TAAGCATTTTAGACTTTTTTTT 18 STAT3_NM_003150 CCTTGCTGACATCCAAATAG 858 CATTGCACTTTTTAACCTTGCTGA 878 _human_2987 CATCCAAATAGAAGATAGGAC 19 STAT3_NM_003150 AAATTAAGAAATAATAACAA 859 CCTAGGTTTCTTTTTAAATTAAGA 879 _human_3041 AATAATAACAATTAAAGGGCA 20 STAT3_NM_003150 TTTTAAATTAAGAAATAATA 860 AAGCCCTAGGTTTCTTTTTAAATT 880 _human_3037 AAGAAATAATAACAATTAAAG Accession: NM_006290 HUGO gene symbol: TNFAIP3 1 TNFAIP3_NM_006290 AGCTTGAACTGAGGAGTAAA 881 ACTTCTAAAGAAGTTAGCTTGAAC 901 _human_3451 TGAGGAGTAAAAGTGTGTACA 2 TNFAIP3_NM_006290 CCTTTGCAACATCCTCAGAA 882 AATACACATATTTGTCCTTTGCAA 902 _human_916 CATCCTCAGAAGGCCAATCAT 3 TNFAIP3_NM_006290 TTCTTTCCAAAGATACCAAA 883 ACGAATCTTTATAATTTCTTTCCAA 903 _human_4422 AGATACCAAATAAACTTCAG 4 TNFAIP3_NM_006290 TTATTTTATTACAAACTTCA 884 TGTAATTCACTTTATTTATTTTATT 904 _human_3688 ACAAACTTCAAGATTATTTA 5 TNFAIP3_NM_006290 TATTTATACTTATTATAAAA 885 GTGAAAAAAAGTAATTATTTATAC 905 _human_4536 TTATTATAAAAAGTATTTGAA 6 TNFAIP3_NM_006290 CATTTCAGACAAAATGCTAA 886 AAGGCCAATCATTGTCATTTCAGA 906 _human_949 CAAAATGCTAAGAAGTTTGGA 7 TNFAIP3_NM_006290 ATGAAGGAGAAGCTCTTAAA 887 GATCCTGAAAATGAGATGAAGGA 907 _human_1214 GAAGCTCTTAAAAGAGTACTTA 8 TNFAIP3_NM_006290 ATTTTGTGTTGATCATTATT 888 AGTTGATATCTTAATATTTTGTGT 908 _human_4489 TGATCATTATTTCCATTCTTA 9 TNFAIP3_NM_006290 TTCATCGAGTACAGAGAAAA 889 TTTTGCACACTGTGTTTCATCGAG 909 _human_2204 TACAGAGAAAACAAACATTTT 10 TNFAIP3_NM_006290 TTACTGGGAAGACGTGTAAC 890 AAAAATTAGAATATTTTACTGGGA 910 _human_3394 AGACGTGTAACTCTTTGGGTT 11 TNFAIP3_NM_006290 TCATTGAAGCTCAGAATCAG 891 ACTGCCAGAAGTGTTTCATTGAA 911 _human_2355 GCTCAGAATCAGAGATTTCATG 12 TNFAIP3_NM_006290 TTCCATTCTTAATGTGAAAA 892 TGTGTTGATCATTATTTCCATTCTT 912 _human_4508 AATGTGAAAAAAAGTAATTA 13 TNFAIP3_NM_006290 TGAAGGATACTGCCAGAAGT 893 TGGAAGCACCATGTTTGAAGGAT 913 _human_2332 ACTGCCAGAAGTGTTTCATTGA 14 TNFAIP3_NM_006290 CACAAGAGTCAACATTAAAA 894 ATAAATGTAACTTTTCACAAGAGT 914 _human_4650 CAACATTAAAAAATAAATTAT 15 TNFAIP3_NM_006290 AATTATTTATACTTATTATA 895 AATGTGAAAAAAAGTAATTATTTA 915 _human_4533 TACTTATTATAAAAAGTATTT 16 TNFAIP3_NM_006290 TTCGTGCTTCTCCTTATGAA 896 CATATTCATCGATGTTTCGTGCTT 916 _human_3907 CTCCTTATGAAACTCCAGCTA 17 TNFAIP3_NM_006290 TATTTTATTACAAACTTCAA 897 GTAATTCACTTTATTTATTTTATTA 917 _human_3689 CAAACTTCAAGATTATTTAA 18 TNFAIP3_NM_006290 TATTACAAACTTCAAGATTA 898 TCACTTTATTTATTTTATTACAAAC 918 _human_3694 TTCAAGATTATTTAAGTGAA 19 TNFAIP3_NM_006290 CTCTTAAAGTTGATATCTTA 899 TGTTTTCATCTAATTCTCTTAAAGT 919 _human_4467 TGATATCTTAATATTTTGTG 20 TNFAIP3_NM_006290 TTCCAAAGATACCAAATAAA 900 ATCTTTATAATTTCTTTCCAAAGAT 920 _human_4426 ACCAAATAAACTTCAGTGTT Accession: NM_003326 HUGO gene symbol: TNFSF4 1 TNFSF4_NM_003326 AATTTGACTTAGCCACTAAC 921 GAGATCAGAATTTTAAATTTGACT 941 _human_2984 TAGCCACTAACTAGCCATGTA 2 TNFSF4_NM_003326 GATATTAATAATATAGTTAA 922 GAGAGTATTAATATTGATATTAAT 942 _human_3422 AATATAGTTAATAGTAATATT 3 TNFSF4_NM_003326 CTGTGAATGCACATATTAAA 923 TGCTTACAGTGTTATCTGTGAATG 943 _human_3119 CACATATTAAATGTCTATGTT 4 TNFSF4_NM_003326 GTTTTCTATTTCCTCTTAAG 924 GGATTTTTTTTTCCTGTTTTCTATT 944 _human_2208 TCCTCTTAAGTACACCTTCA 5 TNFSF4_NM_003326 AAATAGCACTAAGAAGTTAT 925 ATTCAATCTGATGTCAAATAGCAC 945 _human_1727 TAAGAAGTTATTGTGCCTTAT 6 TNFSF4_NM_003326 CCAATCCCGATCCAAATCAT 926 AATGCTTAAGGGATTCCAATCCC 946 _human_3311 GATCCAAATCATAATTTGTTCT 7 TNFSF4_NM_003326 CTATTTAGAGAATGCTTAAG 927 TTAGTTAGATATTTTCTATTTAGA 947 _human_3286 GAATGCTTAAGGGATTCCAAT 8 TNFSF4_NM_003326 CAGTTTGCATATTGCCTAAA 928 AGGTTAAATTGATTGCAGTTTGCA 948 _human_1222 TATTGCCTAAATTTAAACTTT 9 TNFSF4_NM_003326 CTCGAATTCAAAGTATCAAA 929 TATCACATCGGTATCCTCGAATTC 949 _human_326 AAAGTATCAAAGTACAATTTA 10 TNFSF4_NM_003326 ATCTGTGAATGCACATATTA 930 TATGCTTACAGTGTTATCTGTGAA 950 _human_3117 TGCACATATTAAATGTCTATG 11 TNFSF4_NM_003326 TTTGTGGGAAAAGAATTGAA 931 TATACATGGCAGAGTTTTGTGGG 951 _human_2938 AAAAGAATTGAATGAAAAGTCA 12 TNFSF4_NM_003326 ATTGACCATGTTCTGCAAAA 932 ATTTCACTTTTTGTTATTGACCATG 952 _human_2537 TTCTGCAAAATTGCAGTTAC 13 TNFSF4_NM_003326 GATTCTTCATTGCAAGTGAA 933 GGTGGACAGGGCATGGATTCTTC 953 _human_776 ATTGCAAGTGAAGGAGCCTCCC 14 TNFSF4_NM_003326 GATGTCAAATAGCACTAAGA 934 TATCAAATTCAATCTGATGTCAAA 954 _human_1721 TAGCACTAAGAAGTTATTGTG 15 TNFSF4_NM_003326 GTATACAGGGAGAGTGAGAT 935 AAGAGAGATTTTCTTGTATACAG 955 _human_1459 GGAGAGTGAGATAACTTATTGT 16 TNFSF4_NM_003326 GTTGCTATGAGTCAAGGAGT 936 AATGTCTATGTTCTTGTTGCTATG 956 _human_3152 AGTCAAGGAGTGTAACCTTCT 17 TNFSF4_NM_003326 TAGTTGAAATGTCCCCTTAA 937 GTATCCCCTTATGTTTAGTTGAAA 957 _human_1882 TGTCCCCTTAACTTGATATAA 18 TNFSF4_NM_003326 CTCTGTGCCAAACCTTTTAT 938 GATGATTTGTAACTTCTCTGTGCC 958 _human_1980 AAACCTTTTATAAACATAAAT 19 TNFSF4_NM_003326 CTCTGTCTAGAAATACCATA 939 ATGAAAAATAATGATCTCTGTCTA 959 _human_1770 GAAATACCATAGACCATATAT 20 TNFSF4_NM_003326 GGTTTCAAGAAATGAGGTGA 940 CACAGAAACATTGCTGGTTTCAA 960 _human_1680 GAAATGAGGTGATCCTATTATC Accession: NM_006293 HUGO gene symbol: TYRO3 1 TYRO3_NM_006293 AGTTGCTGTTTAAAATAGAA 961 CATTTCCAAGCTGTTAGTTGCTGTT 981 _human_3927 TAAAATAGAAATAAAATTGA 2 TYRO3_NM_006293 CTGTTTAAAATAGAAATAAA 962 CCAAGCTGTTAGTTGCTGTTTAAAA 982 _human_3932 TAGAAATAAAATTGAAGACT 3 TYRO3_NM_006293 GGCATCAGCGATGAACTAAA 963 ACATTGGACAGCTTGGGCATCAGC 983 _human_1731 GATGAACTAAAGGAAAAACTG 4 TYRO3_NM_006293 AATATCCTAAGACTAACAAA 964 GCTACCAAATCTCAAAATATCCTAA 984 _human_3699 GACTAACAAAGGCAGCTGTG 5 TYRO3_NM_006293 GTTGCTGTTTAAAATAGAAA 965 ATTTCCAAGCTGTTAGTTGCTGTTT 985 _human_3928 AAAATAGAAATAAAATTGAA 6 TYRO3_NM_006293 AAAATAGAAATAAAATTGAA 966 TGTTAGTTGCTGTTTAAAATAGAAA 986 _human_3938 TAAAATTGAAGACTAAAGAC 7 TYRO3_NM_006293 CTGTGAAGCTCACAACCTAA 967 GAGCACCATGTTTTCCTGTGAAGC 987 _human_842 TCACAACCTAAAAGGCCTGGC 8 TYRO3_NM_006293 TTGAAGACTAAAGACCTAAA 968 AAAATAGAAATAAAATTGAAGACT 988 _human_3953 AAAGACCTAAAAAAAAAAAAA 9 TYRO3_NM_006293 TCCTAAGACTAACAAAGGCA 969 CCAAATCTCAAAATATCCTAAGACT 989 _human_3703 AACAAAGGCAGCTGTGTCTG 10 TYRO3_NM_006293 GGACATTTCCAAGCTGTTAG 970 GGTCCTAGCTGTTAGGGACATTTC 990 _human_3909 CAAGCTGTTAGTTGCTGTTTA 11 TYRO3_NM_006293 ATGTTTCCATGGTTACCATG 971 AGGAGTGGGGTGGTTATGTTTCCA 991 _human_3190 TGGTTACCATGGGTGTGGATG 12 TYRO3_NM_006293 TAGTTGCTGTTTAAAATAGA 972 ACATTTCCAAGCTGTTAGTTGCTGT 992 _human_3926 TTAAAATAGAAATAAAATTG 13 TYRO3_NM_006293 AAAATTGAAGACTAAAGACC 973 GTTTAAAATAGAAATAAAATTGAA 993 _human_3949 GACTAAAGACCTAAAAAAAAA 14 TYRO3_NM_006293 AGCTGTTAGGGACATTTCCA 974 CATGGGGCGGGTCCTAGCTGTTAG 994 _human_3900 GGACATTTCCAAGCTGTTAGT 15 TYRO3_NM_006293 GAGGACGTGTATGATCTCAT 975 CCTCCGGAGTGTATGGAGGACGTG 995 _human_2511 TATGATCTCATGTACCAGTGC 16 TYRO3_NM_006293 TTTTAGGTGAGGGTTGGTAA 976 CCTTGTAATATTCCCTTTTAGGTGA 996 _human_3400 GGGTTGGTAAGGGGTTGGTA 17 TYRO3_NM_006293 AGCTGACATCATTGCCTCAA 977 TGTGAAGATGCTGAAAGCTGACAT 997 _human_1895 CATTGCCTCAAGCGACATTGA 18 TYRO3_NM_006293 AAATCTCAAAATATCCTAAG 978 TCTGAGCACGCTACCAAATCTCAA 998 _human_3690 AATATCCTAAGACTAACAAAG 19 TYRO3_NM_006293 AAGCTGTTAGTTGCTGTTTA 979 GTTAGGGACATTTCCAAGCTGTTA 999 _human_3919 GTTGCTGTTTAAAATAGAAAT 20 TYRO3_NM_006293 TCCTTGTAATATTCCCTTTT 980 AGTCACAAAGAGATGTCCTTGTAA 1000 _human_3384 TATTCCCTTTTAGGTGAGGGT Accession: NM_000546 HUGO gene symbol: TP53 1 TP53_NM_000546_ TGTTTGGGAGATGTAAGAAA 81 TTTTACTGTGAGGGATGTTTGGG 101 human_1630 AGATGTAAGAAATGTTCTTGCA 2 TP53_NM_000546_ GCATTGTGAGGGTTAATGAA 82 CCTACCTCACAGAGTGCATTGTGA 102 human_1808 GGGTTAATGAAATAATGTACA 3 TP53_NM_000546_ TCGATCTCTTATTTTACAAT 83 TATCCCATTTTTATATCGATCTCTT 103 human_2538 ATTTTACAATAAAACTTTGC 4 TP53_NM_000546_ TGTGAGGGTTAATGAAATAA 84 CCTCACAGAGTGCATTGTGAGGG 104 human_1812 TTAATGAAATAATGTACATCTG 5 TP53_NM_000546_ GAGTATTTGGATGACAGAAA 85 GGAAATTTGCGTGTGGAGTATTT 105 human_812 GGATGACAGAAACACTTTTCGA 6 TP53_NM_000546_ GGATGTTTGGGAGATGTAAG 86 GGTTTTTACTGTGAGGGATGTTTG 106 human_1627 GGAGATGTAAGAAATGTTCTT 7 TP53_NM_000546_ GAAATGTTCTTGCAGTTAAG 87 GTTTGGGAGATGTAAGAAATGTT 107 human_1646 CTTGCAGTTAAGGGTTAGTTTA 8 TP53_NM_000546_ ATGTACATCTGGCCTTGAAA 88 AGGGTTAATGAAATAATGTACAT 108 human_1831 CTGGCCTTGAAACCACCTTTTA 9 TP53_NM_000546_ AGAAATGTTCTTGCAGTTAA 89 TGTTTGGGAGATGTAAGAAATGT 109 human_1645 TCTTGCAGTTAAGGGTTAGTTT 10 TP53_NM_000546_ GGTGAACCTTAGTACCTAAA 90 GTCTGACAACCTCTTGGTGAACCT 110 human_2015 TAGTACCTAAAAGGAAATCTC 11 TP53_NM_000546_ TAACTTCAAGGCCCATATCT 91 CTGTTGAATTTTCTCTAACTTCAA 111 human_1753 GGCCCATATCTGTGAAATGCT 12 TP53_NM_000546_ CTTATCCGAGTGGAAGGAAA 92 GCCCCTCCTCAGCATCTTATCCGA 112 human_782 GTGGAAGGAAATTTGCGTGTG 13 TP53_NM_000546_ ATGATCTGGATCCACCAAGA 93 CATCTCTTGTATATGATGATCTGG 113 human_2086 ATCCACCAAGACTTGTTTTAT 14 TP53_NM_000546_ AATTTTCTCTAACTTCAAGG 94 TGTCCCTCACTGTTGAATTTTCTCT 114 human_1744 AACTTCAAGGCCCATATCTG 15 TP53_NM_000546_ TCTCTTATTTTACAATAAAA 95 CCATTTTTATATCGATCTCTTATTT 115 human_2542 TACAATAAAACTTTGCTGCC 16 TP53_NM_000546_ TTATTTTACAATAAAACTTT 96 TTTTATATCGATCTCTTATTTTACA 116 human_2546 ATAAAACTTTGCTGCCACCT 17 TP53_NM_000546_ GCCTTGAAACCACCTTTTAT 97 AATAATGTACATCTGGCCTTGAAA 117 human_1842 CCACCTTTTATTACATGGGGT 18 TP53_NM_000546_ TATATCGATCTCTTATTTTA 98 TTTATATCCCATTTTTATATCGATC 118 human_2534 TCTTATTTTACAATAAAACT 19 TP53_NM_000546_ CCTTAGTACCTAAAAGGAAA 99 CAACCTCTTGGTGAACCTTAGTAC 119 human_2021 CTAAAAGGAAATCTCACCCCA 20 TP53_NM_000546_ CATTGTGAGGGTTAATGAAA 100 CTACCTCACAGAGTGCATTGTGA 120 human_1809 GGGTTAATGAAATAATGTACAT 

1. An immune modulator comprising an sdRNA, wherein the sdRNA comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification, and wherein the sdRNA is capable of suppressing expression of HAVCR2.
 2. An immunogenic composition comprising an sdRNA, wherein the sdRNA comprises a guide strand and a passenger strand, wherein the guide strand is about 19-25 nucleotides long, and the passenger strand is about 10-19 nucleotides long, wherein the sdRNA includes a double stranded region and a single stranded region, wherein the single stranded region includes 5-9 phosphorothioate modifications, wherein the sdRNA is chemically modified, including at least one 2′-O-methyl modification or 2′-fluoro modification, wherein the sdRNA is capable of suppressing expression of HAVCR2, and wherein the immunogenic composition further comprises immune cells modified by the sdRNA to suppress expression of HAVCR2.
 3. The immunogenic composition of claim 2, wherein the immune cells within the composition are further modified to suppress expression of a plurality of immune checkpoint genes.
 4. The immunogenic composition of claim 2, wherein said cells are modified to suppress expression of at least one immune checkpoint gene, and at least one gene selected from the group consisting of: an anti-apoptosis gene, a cytokine receptor gene, and a regulator gene. 5-8. (canceled)
 9. The immunogenic composition of claim 3, wherein said modified cells further comprise one or more sdRNA capable of suppressing expression of at least one target gene selected from the group consisting of: CTLA4, PD1/PDCD1, TGFBR1, TGFRBR2, IL10RA, TP53, BAX, BAK1, CASP8, ADORA2A, LAG3, CCL17, CCL22, DLL2, FASLG, CD274, IDO1, ILIORA, JAG1, JAG2, MAPK14, SOCS1, STAT3, TNFAIP3, TYRO3, BTLA, KIR, B7-H3 receptor, and B7-H4 receptor.
 10. The immunogenic composition of claim 9, wherein said cells are selected from the group consisting of T-cells, NK-cells, antigen-presenting cells, dendritic cells, stem cells, induced pluripotent stem cells, and/or stem central memory T-cells.
 11. The immunogenic composition of claim 9, wherein said cells are T-cells comprising one or more transgene expressing high affinity T-cell receptors (TCR) and/or chimeric antibody-T-cell receptors (CAR).
 12. (canceled)
 13. The immunogenic composition of claim 9, wherein said cells comprise one or more sdRNA targeting one or more anti-apoptotic genes selected from the group consisting of BAX, BAC, TP53, and Casp8.
 14. (canceled)
 15. The immunogenic composition of claim 9, wherein said sdRNA comprises at least one 2′-O-methyl modification and/or at least one 2′-O-Fluoro modification, and at least one phosphorothioate modification.
 16. The immunogenic composition of claim 9, wherein said sdRNA comprises at least one hydrophobic modification.
 17. The immunogenic composition of claim 9, wherein said sdRNA is modified to comprises at least one cholesterol molecule.
 18. The immunogenic composition of claim 9, wherein said sdRNA targets a sequence selected from SEQ ID NOs: 1-360 and 381-1000.
 19. A method of producing the immunogenic composition of claim 3, said method comprising transforming at least one cell with one or more sdRNA capable of targeting and suppressing expression of one or more immune target genes, wherein at least one sdRNA interferes with HAVCR2 expression.
 20. The method of claim 19, wherein the at least one cell comprises an sdRNA that inhibits expression of one or more genes selected from the group consisting of: CTLA4, PD1/PDCD1, TGFBR1, TGFRBR2, IL10RA, TP53, BAX, BAK1, CASP8, ADOARA2A, LAG3, CCL17, CCL22, DLL2, FASLG, CD274, IDO1, ILIORA, JAG1, JAG2, MAPK14, SOCS1, STAT3, TNFAIP3, TYRO3, BTLA, KIR, B7-H3 receptor, and B7-H4 receptor.
 21. The method of claim 19, wherein said cells are selected from the group consisting of T-cells, NK-cells, antigen-presenting cells, dendritic cells, stem cells, induced pluripotent stem cells, and/or stem central memory T-cells. 22-24. (canceled)
 25. A method for treating a subject having proliferative disease or infectious disease, the method comprising administering to the subject the immune modulator of claim
 1. 26. The method of claim 25, wherein said proliferative disease is cancer.
 27. The method of claim 25, wherein said infectious disease is a pathogen infection.
 28. (canceled)
 29. The immune modulator of claim 1, wherein at least one sdRNA targets a sequence selected from SEQ ID NOs: 361-380.
 30. The immunogenic composition of claim 2, wherein at least one sdRNA targets a sequence selected from SEQ ID NOs: 361-380. 